Review

Free Access

Aegerolysins: Structure, function, and putative biological role

Sabina Berne

Medical Centre for Molecular Biology, Institute of Biochemistry, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia

Search for more papers by this author

Ljerka Lah,

Ljerka Lah

Ljerka Lah, Laboratory for Biosynthesis and Biotransformation, National Institute of Chemistry, 1000 Ljubljana, Slovenia

Search for more papers by this author

Kristina Sepčić,

Corresponding Author

Kristina Sepčić

[email protected]

Department of Biology, Biotechnical Faculty, University of Ljubljana, 1000 Ljubljana, Slovenia

University of Ljubljana, Večna pot 111, 1000 Ljubljana, SloveniaSearch for more papers by this author

Sabina Berne,

Sabina Berne

Medical Centre for Molecular Biology, Institute of Biochemistry, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia

Search for more papers by this author

Ljerka Lah,

Ljerka Lah

Ljerka Lah, Laboratory for Biosynthesis and Biotransformation, National Institute of Chemistry, 1000 Ljubljana, Slovenia

Search for more papers by this author

Kristina Sepčić,

Corresponding Author

Kristina Sepčić

[email protected]

Department of Biology, Biotechnical Faculty, University of Ljubljana, 1000 Ljubljana, Slovenia

University of Ljubljana, Večna pot 111, 1000 Ljubljana, SloveniaSearch for more papers by this author

First published: 12 February 2009

https://doi.org/10.1002/pro.85

Citations: 68

Share a link

Email
Wechat
Bluesky

Abstract

Aegerolysins, discovered in fungi, bacteria and plants, are highly similar proteins with interesting biological properties. Certain aegerolysins possess antitumoral, antiproliferative, and antibacterial activities. Further possible medicinal applications include their use in the prevention of atherosclerosis, or as vaccines. Additional biotechnological value of fungal aegerolysins lies in their involvement in development, which could improve cultivation of commercially important edible mushrooms. Besides, new insights on microheterogeneity of raft-like membrane domains could be gained by using aegerolysins as specific markers in cell and molecular biology. Although the exact function of aegerolysins in their producing organisms remains to be explained, they are biochemically well characterized all-β structured proteins sharing the following common features: low isoelectric points, similar molecular weights (15–17 kDa), and stability in a wide pH range.

Introduction

In 2002, a new aegerolysin protein family (PF06355; IPR 009413) was defined, comprised of a few isolated and sequenced proteins, and several genomic transcripts and sequences predicted from expressed sequence tags. The first aegerolysin-like protein to be isolated1 and sequenced2 was Asp-hemolysin, a lethal and cardiotoxic component of the pathogenic fungus Aspergillus fumigatus. This opportunistic pathogen causes a wide spectrum of serious diseases including invasive aspergillosis. The pathogeneicity of A. fumigatus is a consequence of action of a series of toxic substances (cell wall components, allergens, pigments, adhesins, enzymes and toxins) which seem to act in an additive and/or synergic way on the cells.3 Asp-hemolysin was proposed to contribute to A. fumigatus pathogenesis.3 After its purification in the early seventies, a lot of work has been done to determine its structure and biological effects.

Subsequently, with the beginning of the genomics era, several other similar proteins were discovered. The first protein showing considerable amino acid sequence identity (43%) with Asp-hemolysin was the transcript of the Aa-Pri1 gene from the edible black poplar mushroom Agrocybe aegerita,4 which was soon followed by two putative hemolysin-like proteins from the bacterium Clostridium bifermentans.5 During the last few years, several other proteins or cDNA transcripts with highly identical sequences have been discovered6-11 and assigned to aegerolysins (Supporting Information). The isolated proteins and their recombinant forms were found to be acidic, to have a molecular weight from 15 to 17 kDa, and to share some important biological properties like membrane activity and a putative role in development.

The aim of this review is to summarize the current knowledge on this novel protein family, and to discuss the structure, function and possible biological role of aegerolysins.

Purification of Native and Recombinant Aegerolysin-Like Proteins

Much effort was invested in purification of the first aegerolysin-like protein, Asp-hemolysin from A. fumigatus. As this pathogenic mould produces various putative virulence factors, the purification was accomplished by monitoring the hemolytic activity during the isolation procedure, and separating the hemolytic compound from other toxic (e.g., dermonecrotic, neurotoxic, and nephrotoxic) molecules. Indications of a proteinaceous nature of the A. fumigatus hemolytic component can already be found in a very early report,12 which describes the loss of half of its toxicity after heating. This observation was later corroborated by other authors who had tried to purify the hemolytic compound(s) from the centrifuged mycelial homogenate.13, 14 The purification procedure was further improved by precipitation of the hemolytic compound with acetone and ammonium sulfate,14, 15 and column electrophoresis and ion-exchange chromatography on DEAE-cellulose.16 The protein was finally purified to homogeneity from the Fresenius-Muramatsu A. fumigatus strain, isolated from the lung of an infected penguin.1 The purification included precipitation with ammonium sulfate, anionic column chromatography and additional gel filtrations. This basic scheme was later used for purification of all other aegerolysins.7, 9, 17 In 2006, Ngai and Ng have simplified the purification procedure by gel filtration of crude fungal extract on FPLC, followed by ion-exchange chromatography and second gel filtration.

There are some peculiarities concerning the isolation of fungal aegerolysins. Ostreolysin and pleurotolysin A, for instance, can be isolated in 10-fold higher yields from freshly collected oyster mushrooms (Pleurotus ostreatus) than from frozen ones9(Berne, personal communication). Optimal isolation is achieved from primordia and young fruiting bodies of P. ostreatus, which coincides with the observation that the mushroom contains the highest concentration of hemolysin during this developmental stage.7, 18

Several expression constructs were designed to produce recombinant aegerolysins. Recombinant Asp-hemolysin was expressed in Escherichia coli as a fusion protein with maltose-binding protein (MBP) allowing a rapid, one-step purification on amylose resin.19 The purified protein exhibited the same immunoreactivity with anti-Asp-hemolysin antibodies as native Asp-hemolysin, and was able to bind the oxidized form of low-density plasma lipoprotein (ox-LDL), but did not show any hemolytic activity observed with the native protein. Efficient expression of Clostridium bifermentans cbm17.1 and cbm17.2 genes was achieved by introducing them as His₆-tagged proteins in E. coli and Bacillus thuringiensis.5 However, when recombinant proteins Cbm17.1 and Cbm17.2 were assayed for biological activity, no toxicity to mosquito larvae was observed and no hemolytic action recorded.20 The His₆-tagged PA0122 protein from Pseudomonas aeruginosa was also successfully expressed in E. coli.11 Purified recombinant PA0122 (rPA0122) protein was used for immunization and binding assays. It specifically bound to ox-LDL and lysophosphatidylcholine with an affinity equivalent to recombinant Asp-hemolysin, but similarly, did not exhibit any hemolytic activity.

In contrast to other aegerolysins, pleurotolysin is a two-component sphingomyelin-specific cytolysin.9 Recombinant pleurotolysin A (rPlyA) was expressed in E. coli as a soluble protein with the same molecular mass (17 kDa) and identical N-terminal amino acid sequence as natural PlyA. Recombinant pleurotolysin B (rPlyB) was obtained as a functional 59-kDa protein by unfolding and refolding of the insoluble E. coli fraction. In addition, recombinant PlyA and Ply B were immunostained with specific antisera against PlyA and PlyB, respectively. As each component lacked hemolytic activity in the absence of its counterpart, they were combined to study pleurotolysin pore-forming properties.21

Characterization of Aegerolysin-Like Proteins

The aegerolysin protein domain

In the Pfam protein database22 there are currently 43 members with the aegerolysin protein domain (PF06355) from 21 different species. Our PSI-BLAST23, 24 search (July, 2008) of the nonredundant protein database at NCBI (http://www.ncbi.nlm.nih.gov/BLAST), using the Aa-Pri1 translated amino acid sequence (O42717) from A. aegerita as query confirmed the Pfam entries. Additional fungal and plant homologs were retrieved from the expressed sequence tags (EST) database at NCBI, and from the Fungal Genome Initiative (FGI) database, Broad Institute (http://www.broad.mit.edu/annotation/fungi/fgi/) using the same query and the tblastn algorithm. In constructing our aegerolysin protein alignment (see Fig. 1), highly similar strain-specific sequences (e.g., 98% redundancy in Pseudomonas aeruginosa), and sequence fragments (e.g., in Bacillus thuringiensis) were omitted. Utilizing the consensus sequence, generated by Jalview,28 aegerolysin secondary structure was predicted by Jpred3,29 hyprospII,30 and PredictProtein.31 All three prediction tools generated almost identical results, proposing aegerolysins as all-beta proteins. This structural design was confirmed experimentally in the case of ostreolysin. CD spectroscopy revealed that this aegerolysin-like protein at pH 7.8 has 66% β-structure (6% β-turn and 60% β-sheet), 10% α-helix and the rest aperiodic secondary structure.32 These data agree with FTIR spectroscopy results.33

Details are in the caption following the image — **Figure 1**
Open in figure viewer PowerPoint

Protein alignment of the aegerolysin family members. Sequences were retrieved from Entrez (http://www.ncbi.nlm.nih.gov/sites/gquery) and manually edited to include only the aegerolysin domain for an optimal alignment generated by M-Coffee.25, 26 The alignment output was edited using GeneDoc27 conserved mode of shading, showing percentage of conservation (60%, light gray; 80%, grey; 100%, dark grey) within a column in the alignment. Consensus (boxed in red) generated by Jalview28 was used to predicit aegerolysin secondary structure. The distribution of secondary structure elements is presented bellow the protein alignment (green box-extended strand, grey line - random coil). The accession numbers of the proteins in Figure 1 are as follows: aegerolysin (gi 24636240), ostreolysin (gi 60461919), pleurotolysin_A (gi 54312022), T_versicolor (gi 90639437), H_annosum (gi 186897694), Asp-hemolysin (gi 70985747), A_clavatus (gi 121709507), A_nidulans (gi 67522192), A_niger_1 (gi 145230219), A_niger_2 (gi 145256342), A_oryzae_1 (gi 169777319), A_oryzae_2 (gi 169772307), A_oryzae_3 (gi 169785219), A_terreus (gi 115390458), N_fischeri (gi 119487614), B_cinerea (gi 116087898), C_immitis (gi 119182290), P_brasiliensis (gi 46346063), N_crassa (gi 85079504), B_vulgaris (gi 26112720), L_multiflorum (gi 46507636), R_australe (gi 158524422), P_aeruginosa_1 (gi 152985646), P_aeruginosa_2 (gi 15595320), C_bifermentans_1 (gi 2292821), C_bifermentans_2 (gi 2292820), B_thuringiensis_1 (gi 14571542), B_thuringiensis_2 (gi 49175503).

Aegerolysin protein domain, derived from the multiple sequence alignment and hidden Markov models (HMMs), is rich in aromatic (around 10%) and charged (20–24%) amino acid residues (pepstats predicition).34 Basidiomycete and Aspergillus aegerolysins contain a cluster of negatively charged amino acids (see Fig. 2). This segment, homologous to the ApoB-binding site of human low-density-lipoprotein receptor,35 was demonstrated to bind ox-LDL, and lysophosphatidycholine as a typical lipid moiety of ox-LDL.11, 36, 37 Furthermore, as proposed by Sakurai et al.,21 this negatively charged region may be involved in the recognition of choline moiety of sphingomyelin. Another feature of aegerolysin protein domain is conservation of cysteine and certain trypthophan residues (see Fig. 2) implying their possible structural and/or functional roles. Intact cysteine residues have been proven indispensable for hemolytic activity of ostreolysin7 and Asp-hemolysin.38 Tryptophan-rich regions have been reported in some other pore-forming proteins as responsible for the initial attachment to the membrane.39, 40 Moreover, combined aromatic residues and a cysteine residue have been suggested to be involved in sterol binding of cholesterol-dependent bacterial pore-forming toxins.41

Taxonomic distribution of aegerolysin-like proteins

Aegerolysin-like proteins appear in eukaryotic and bacterial taxa (Fig. 3). Most species (18) with genes encoding aegerolysins are found within Fungi, and belong to two sister groups, the Ascomycetes and the Basidiomycetes. In the former, the Aspergillus genus has the highest number od species (8) with aegerolysin genes. All fungal species with aegerolysin genes are either filamentous or dimorphic. Interestingly, the only other eukaryotic group with genes encoding aegerolysin-like proteins is Plantae. Blast searches revealed two eudicotyledonous and one monocotyledonous species to contain aegerolysin ESTs. Aegerolysin-like proteins also appear in two different taxonomic groups in Bacteria; the Firmicutes and the gamma-proteobacteria. To further diversify the distribution of aegerolysin-like proteins, there is also one viral representative, the Trichoplusia ni ascovirus 2c.

The evolutionary relationships between the relatively few members of the aegerolysin protein family are difficult to establish and the phylogenetic tree is left unresolved (tree not shown). Well-supported phylogenetic relationships can be discerned only among the smaller taxa, for example, in Basidiomycetes, or the numerous strain-specific homologs in Bacillus thuringiensis. The latter is perhaps a case of microheterogeneity, where numerous aegerolysin-like homologs constitute a set of structurally similar, yet functionally diverse proteins.45

As to the origin of the aegerolysin-encoding gene, we can hypothesize that it first appeared in Fungi, in the last common ancestor of Ascomycetes and Basidiomycetes, where it is present in the greatest number of species. This assumption could be supported by the putative biological role of these proteins in fruiting body development, signaling, and their role as potential virulence factors in pathogenic species. There were only three species of the numerous sequenced bacterial genomes found to contain aegerolysin-encoding genes, belonging to two divergent taxa. A readily available explanation for bacterial aegerolysin family members is horizontal gene transfer (HGT) from fungi. Fluorescent pseudomonads were found to be closely associated with the cultivated mushroom Pleurotus ostreatus,46 which is a rich source of aegerolysin-like proteins.7, 9 The incorporation of aegerolysin homologs into bacterial genomes could be beneficial in the same respects as their putative biological roles are beneficial to fungi.

More puzzling is the appearance of aegerolysins in plants, again found in only three species belonging to monocots and dicots, in the EST database. This might also be a case of HGT, although reports of HGT to plant nuclear DNA are rare.47 HGT could have had occurred either from fungi or bacteria, which both form an intimate relationship with the host organism as plant pathogens or symbionts. Such close ecological associations could provide opportunities for gene transfer events.48 It could, however, also be a result of plant samples contaminated with mould (e.g., Lolium multiflorum leaf samples infected with powdery mildew [Ikeda, S., 2004 (unpublished), EST sequence database at NCBI)].

Physical and biochemical characterization of aegerolysin-like proteins

Proteins from the aegerolysin family share several common features. They all have very low isoelectric points, similar molecular weights, and are heat-labile (see Table I). Furthermore, they seem to be stable in a wide range of pH.1, 10, 32 In contrast to most of the known aegerolysins, that exert their biological properties (mainly the membrane lytic properties) in their monomeric forms,1, 7, 10, 17 pleurotolysin A isolated from the mushroom Pleurotus ostreatus acts as a bi-component cytolysin in concert with a 59 kDa pleurotolysin B. When coupled, these two proteins assemble into a ring-shaped transmembrane pore and are named pleurotolysin.9, 21

Table I. Main Physical and Biochemical Characteristics of Aegerolysin-Like Proteins

Species	Protein	Prot param (ExPASy Tools)			Exp. Data		Reference
Species	Protein	Amino acids	MW (Da)	pI	MW (Da)	pI	Reference
Agrocybe aegerita	Aegerolysin Aa-Pri1 precursor	145	16,104	5.65	16,093	5.7	4
	Aegerolysin	10			14,898	4.9	50
	cDNA clone Agf19	145	16,087	6.10			Im, Lee and Shim, 2006 (unpublished)
Heterobasidion anosum	cDNA clone 18A12 (2a)	130	14,236	6.07			51
Pleurotus eryngii	Eryngeolysin	40			17,000		10
Pleurotus ostreatus	Ostreolysin	137	14,855	4.82			32
	Ostreolysin	50			17,000	5.0	7
	Ostreolysin				14,975		52
	Pleurotoysin				12,600a	6.4	17
	Pleurotolysin A	138	15,136	5.87			21
	Pleurotolysin A				17,000		9
Trametes versicolor	cDNA clone TverSEQ10942	174	19,205	5.05			Tsang, Storms and Butler, 2006 (Unpublished)
Aspergillus fumigatus	Asp-hemolysin precursor	139	15,199	5.29			53
	Asp-hemolysin				30,000b	4.0	1
	Asp-hemolysin	131	14,275	5.24			2
	Asp hemolysin-like protein	139	15,820	4.54			53
Aspergillus clavatus	Hypothetical protein ACLA_066510	139	15,644	4.57			Nierman, 2006 (Unpublished)
Aspegillus flavus	Conserved hypothetical protein AFL2G_03935	139	15,571	4.61			FGI Broad Institute
Aspegillus flavus	Predicted protein AFL2G_08431	104	11,365	4.55			FGI Broad Institute
Aspergillus nidulans	Hypothetical protein AN1553.2	136	14,864	4.59			Birren et al. 2003 (Unpublished)
Aspergillus niger	Hypothetical protein An01g09980	145	16,259	7.81			54
Aspergillus niger	Hypothetical protein An19g00210	142	16,256	5.20			54
Aspergillus oryzae	Unnamed protein product AO090701000257	139	15,571	4.61			55
	Unnamed protein product AO090023000032	141	16,359	5.10			55
	Unnamed protein product AO090010000018	144	16,294	9.53			55
Aspergillus terreus	Conserved hypothetical protein ATEG_03556	152	17,022	5.24			Birren et al. 2005 (Unpublished)
Neosartorya fischeri	Asp hemolysin-like protein NFIA_030750	139	15,810	4.64			Nierman et al. 2006 (Unpublished)
Botrytis cinerea	cDNA clone 3498_G06	144	15,795	5.47			56
Coccidioides immitis	Hypothetical protein CIMG_06184	182	20,269	6.30			Birren et al. 2005 (Unpublished)
Coccidioides posadasii	Hypothetical protein CPAG_03541.1	134	14,798	5.01			FGI Broad Institute
Neurospora crassa	Hypothetical protein NCU03475	198	21,401	5.05			8
Paracoccidioides brasiliensis	cDNA clone EST008190	127	14,078	3.88			57
Beta vulgaris	cDNA clone 024-006-B13	135	14,875	6.06			58
Lolium multiflorum	cDNA clone SL010G09-5	135	14,691	6.08			Ikeda, 2004 (Unpublished)
Rheum australe	cDNA clone RaSFL 362	116	12,830	4.51			Ghawana et al. 2007 (Unpublished)
Bacillus thuringiensis	Crystal protein ET80	132	14,839	6.02			59
Bacillus thuringiensis	Cry34A-like protein	97	10,614	5.86			60
Clostridium bifermentans malaysia	Hemolysin-like protein Cbm 17.1	153	17,200	5.41			5
Clostridium bifermentans malaysia	Hemolysin-like protein Cbm 17.2	152	17,463	5.82			5
Pseudomonas aeruginosa	Hypothetical protein PA0122	136	14,579	4.65			6, 11
Pseudomonas aeruginosa	Aegerolysin superfamily	121	12,713	8.36			Dodson et al. 2007 (Unpublished)
Trichoplusia ni ascovirus 2c	Hypothetical protein TNAV2c_gp029	222	23,777	6.19			60

a Lower MW presumably due to proteolytic degradation.
b Probably protein aggregation.

Biological Activities of Aegerolysin-Like Proteins

Interaction with lipid membranes

Hemolytic activity

The first isolated aegerolysin, Asp-hemolysin, was proposed to form distinctive pores in erythrocyte membranes and to induce hemolysis by a colloid-osmotic mechanism. An early study using scanning electron microscopy showed that this hemolysin forms rings and patches at the surface of erythrocyte membranes.61 Aggregation of the protein at the membrane surface and inhibition of hemolysis using osmotic protectants of various molecular weights was later proven also for ostreolysin,33 pleurotolysin,9 and eryngeolysin.10 The hydrodynamic diameters of the resulting pores were estimated to be around 4 nm.9, 33 Pleurotolysin-induced lysis was reported to proceed (i) by the initial binding of the 17 kDa sphingomyelin-binding component, pleurotolysin A, to the membrane, (ii) subsequent addition of pleurotolysin B (59 kDa), and (iii) their association into a 700 kDa SDS-stable, ring-shaped structures with outer and inner diameters of 14 and 7 nm, respectively. Most efficient hemolysis occurred when the two components were mixed in a 3:1 molar ratio, implying that they are incorporated in the pore complex in the same molar ratio.9 All aegerolysins aforementioned were hemolytic in the nanomolar range.

The pH-optima for binding of aegerolysins to the erythrocyte membrane and its permeabilization do not overlap. Optimal binding is usually achieved at pH 5–7, as shown for Asp-hemolysin61 and ostreolysin.32 In contrast, the maximal hemolysis is observed at pH 7–8.17, 32 Aegerolysins however retain their membrane activity in a wide pH range from 3.5 to 10.5.1, 10, 32, 61 Binding of ostreolysin to the membrane correlates with an increase of intrinsic tryptophan fluorescence and the α-helical content of the protein, and with aggregation, suggesting that binding to the bilayers evokes further conformational transitions necessary for insertion and pore formation.32, 33

Asp-hemolysin lytic activity can be abolished by certain divalent ions, like Hg²⁺, Cu²⁺, Fe²⁺, and Pb²⁺, and later restored by the use of β-mercaptoethanol or cysteine.1 Similar inhibition of hemolytic activity by micromolar concentrations of Hg²⁺ and Fe²⁺ was also reported for ostreolysin32 and eryngeolysin,10 respectively. Early studies involving chemical modifications of Asp-hemolysin suggested that while cysteines and arginines were necessary for hemolytic activity,38, 62-64 histidine residues were not. The implication of intact cysteine residues in a hemolytic process was later proven also for ostreolysin.7, 32

Varying interaction with erythrocytes from different species is another interesting feature of assorted aegerolysins. For example, Asp-hemoylsin is more lytic to chicken and human erythrocytes than to those of rodents.1 Its activity against frog erythrocytes is 833-fold lower than against chicken erythrocytes.65 A slightly different pattern, showing greater sensitivity of mammalian erythrocytes in comparison to avian ones, was observed for eryngeolysin.10 The membrane activity of pleurotolysin was proposed to correlate with the sphingomyelin content of erythrocyte membranes, exhibiting the following preferences: sheep > ox > goat > man > cat > rabbit > swine > mouse/rat > horse > dog > guinea pig,17 or sheep > human > rabbit > dog > horse.9 In contrast, ostreolysin-induced lysis of sheep, bovine and human erythrocytes33 was similar, but much less potent against canine66 and rodent67 erythrocytes.

Permeabilization of lipid vesicles

The lytic process of many pore-forming proteins starts with the recognition of a distinct membrane component. The binding of eryngeolysin, for example, was found to be inhibited by millimolar concentrations of N-glycosylneuraminic acid.10 The aegerolysins' lytic activity, however, is not altered upon incubation with pure sphingo- or glycerophoypholipids, or cholesterol.33, 65 Similarly, liposomes composed of egg phosphatidylcholine in combination with various other lipids were unable to inhibit ostreolysin-induced hemolysis.33 In contrast, liposomes composed from sheep erythrocyte sphingomyelin and cholesterol in a 1:1 molar ratio successfully inhibited binding and lytic activity of pleurotolysin,17, 9 and ostreolysin.51 As these effects were absent when liposomes were composed of cholesterol in combination with other lipids, it was proposed that pleurotolysin is a specific sphingomyelin-binding protein.9, 17 However, recent work performed on ostreolysin suggests that the specific interaction of this protein (and probably other aegerolysin-like proteins) with sphingomyelin:cholesterol liposomes could be due to its binding to cholesterol-enriched raft-like membrane microdomains. These highly investigated membrane entities were reported to be involved in several important biological functions, such as exocytosis and endocytosis, signal transduction, pathogen entry, and attachment of various ligands,68-70 and to exist in a so-called liquid-ordered phase. Lipid domains in such a physical state are more resistant to solubilization by detergents than the ones in a liquid-disordered phase.71 Hence, lipid rafts are operationally often called detergent-resistant membranes (DRMs).68 Upon preincubation, ostreolysin was found in isolated DRMs of both sphingomyelin:cholesterol vesicles and Chinese Hamster Ovary cells.51 Further, permeabilization of sphingomyelin:cholesterol (1:1) vesicles by ostreolysin occurred only above 30 mol% cholesterol, the concentration at which this sterol induces the formation of a liquid-ordered phase.51, 72, 73 The interaction of ostreolysin with cholesterol-enriched membrane domains can be diminished or disrupted (i) by the addition of mono- and di-unsaturated phosphatidylcholine,51 (ii) by replacing cholesterol with other natural sterols or cholesterol derivatives,73 (iii) by the addition of micromolar concentrations of fatty acids and lysophospholipids,33, 74 and (iv) by the pre-treatment of cell membranes with a cholesterol scavenger, methyl-β-cyclodextrin.74 Ostreolysin did not affect electron paramagnetic resonance spectra of sphingomyelin:cholesterol (1:1) vesicles,51 which would indicate a recruitment of the lipids upon its binding to the membrane, a feature that was proven for other raft-binding cytolysins like the cholera toxin B subunit.75 It seems that this protein, rather than recognizing sphingomyelin, recognizes and binds to specific patterns formed by cholesterol molecules at the surface of biological membranes.

Cholesterol-dependence of ostreolysin resembles that of bacterial thiol-activated (cholesterol-dependent) cytolysins (CDCs), allowing us to conclude that this protein, and probably other aegerolysin members, could be functionally classified as CDCs.73 Ostreolysin cannot bind pure cholesterol,33 but this sterol seems to be crucial for its membrane activity, which is highly cooperative with respect to cholesterol membrane concentration.51, 73 Similarly, it was recently shown that the sole presence of cholesterol is not a prerequisite for the membrane activity of CDCs,41, 75-78 suggesting furthermore that the distribution of cholesterol molecules on the membrane, and the specific patterns that are subsequently formed, could be of crucial importance for the activity of this group of cytolysins. For example, a cholesterol-binding pore-forming Vibrio cholerae cytolysin, whose crystal structure strongly resembles those of CDCs,79 can bind to pure cholesterol and oligomerize with it in solution, but it prefers association with sphingolipid/cholesterol complexes rather than with individual lipid molecules.80

Finally, the discovery that some aegerolysins bind specifically, and with high affinity to the ox-LDL11, 81 has lead to a more detailed research of interaction between the two molecules. The direct binding of lysophosphatidycholine (which is the major component of ox-LDL) to some aegerolysins, Asp-hemolysin and PA0122, was confirmed by ion-exchange chromatography37 and by surface plasmon resonance experiments respectively.11 Although the membrane activity of ostreolysin was strongly inhibited by lysophospholipids and fatty acids,33, 74 the same binding specificity could not be proven. It is suggested rather that lysophospholipids influence the membrane activity of ostreolysin by altering the physical properties of the membrane, for example, by partitioning into liquid-ordered membrane domains.74

Binding to serum lipoproteins

The membrane activity of aegerolysin-like proteins can be inhibited by the addition of serum.33, 65, 82 Indeed, binding to blood plasma components is another distinctive feature of the most studied aegerolysin-like protein, Asp-hemolysin.83 This protein is similar (26.5% identity and 68.8% similarity over a 34 amino acid overlap) to a human LDL receptor precursor2 and interacts with several lipoprotein components of human plasma, especially ox-LDL.35, 36, 84 The binding to ox-LDL is concentration-dependent, and is reduced if positively charged amino acid residues of the hemolysin are blocked.85 Surface plasmon resonance experiments confirmed the specific binding of Asp-hemolysin to ox-LDL with a K_d of 0.63 μg/mL.81 The binding constants of a recombinant aegerolysin from Pseudomonas aeruginosa (rPA0122) to ox-LDL and lysophosphatidylcholine were very similar to those of Asp-hemolysin.11

Recently, a synthetic peptide (P21) derived from the positively charged region of Asp-hemolysin was found to inhibit the ox-LDL-induced macrophage proliferation.86 Further, using synthetic peptides of 4–29 amino acid residues, the specific ox-LDL-binding sequence (YKDG) was identified.87 The YKDG region is important for binding of these peptides to lysophosphatidylcholine, that seems to be responsible for most of the effects of ox-LDL, including induction of macrophage proliferation,88 which has an important role in the development and progression of atherosclerosis.

Cytotoxic activity

The toxicity of aegerolysins to experimental animals is most probably also a consequence of their cytolytic effects. So far, aegerolysins were found to be cytotoxic for different cell lines (Table II). Direct microscopic observation of certain tumoral cell lines exposed to ostreolysin showed typical signs of a colloid-osmotic hemolysis, like swelling, blebbing and degranulation of cells.33 Similar events were noted during real-time monitoring of ostreolysin-induced disruption of fluorescently stained human chondrocytes and osteoblasts using confocal microscopy.91 Cytotoxicity was found to be affected by the same factors as hemolysis, for example, by chemically blocking the amino- and thiol groups, or by the serum.51, 82 Furthermore, macrophages treated with Asp-hemolysin express mRNA transcripts for several cytokines like TNF-alpha, interleukin-1a and 1b.82 Asp-hemolysin-induced mRNA expression of cytokines such as TNF-alpha, IL-1b, IL-6, IL-8, and granulocyte-macrophage colony stimulating factor was also reported for endothelial cells.90 Finally, eryngeolysin possesses antibacterial activity towards Bacillus subtillis and Bacillus megaterium, and inhibits the mitogenic response of murine splenocytes.10

Table II. Cytotoxic Properties of Aegerolysin-Like Proteins on Different Cell Lines

Protein	Cell line	ED₅₀ (μg/mL)	Time of exposure (h)	Reference
Asp-hemolysin	Leukocytes (human)	500	NA	90
	Alveolar macrophages (guinea pig)	60	NA	90
	Peritoneal macrophages (mouse)	25	1	84
	Umbilical vein endothelial cells (human)	100	1	91
Ostreolysin	HT 1080 (fibrosarcoma, human)	10	2	33
	MCF 7 (mammalian tumour, human)	10	2	33
	Chinese Hamster Ovary cells	1	0.25	52
	Human articular chondrocytes	1	1	92
	Fibroblasts (lungs of Chinese hamster, V-79-379A	1.3	1	93
	Endothelial cells (human umbilical vein, HUVEC)	2.2	1	93
Eryngeolysin	L1210 leukemia (human)	NA	10	10

NA, not available.

Toxicity for experimental animals

Toxic and lethal effects of aegerolysin-like proteins were most extensively studied in Asp-hemolysin from Aspergillus fumigatus. This opportunistic pathogen has been known for a long time to cause neurotoxic,12, 93 dermonecrotic,94, 95 nephrotoxic,94, 96 complement-inhibitory,97 and hemolytic effects.93, 94 The LD₅₀ of Asp-hemolysin for mice and chicken was determined to be 750 and 350 μg/kg, respectively. This presumably secretory protein2, 98 was also detected in vivo during the experimental infection of mice by A. fumigatus spores.98 Histopathological findings in mice after the i.v. administration of Asp-hemolysin comprise perivascular lesions of various degrees in kidney, heart, liver and brain, increase of capillary permeability, contraction of the ileum and cytotoxicity, accompanied by the appropriate biochemical alteration of sera.89, 99

Although the edible oyster mushroom (Pleurotus ostreatus) is a widely cultivated species, sporadic local intoxications following its ingestion by humans and animals were also recorded. It was suggested that the toxicity is associated with thermolabile proteinaceous molecules.100 Bernheimer and Avigad17 did not observe any obvious signs of intoxication in mice after i.v. application of 320 hemolytic units of pleurotolysin. However, recent investigation of ostreolysin revealed cardiorespiratory and toxic effects in rodents, and their death after an i.v. administration with a LD₅₀ of 1175 μg/kg.67 Cardiotoxic effects are probably due to ostreolysin-induced dose-dependent increase in aortic ring tension, and its cytotoxicity to endothelial cells resulting in hyperkalaemia.92 The histopathological effects of ostreolysin in mice were not studied yet. Pleurotus ostreatus water extract, however, was found to provoke effects similar to those induced by Asp-hemolysin: hemorrhages in the intestine, liver, lung and kidney, and degenerative changes in the liver, with a LD₅₀ higher than 3000 μg/kg.100 Such an extract also induced dose-dependent contractions of nonsensitized guinea pig trachea smooth muscle.101

Role of aegerolysins in development

Experimental results imply that aegerolysins play an important role in the growth cycle of the organism that produces them. Fernandez-Espinar and Labarère4 determined that Aa-Pri1 mRNA expression coincided with the developmental stage of primordia, and proposed an involvement of the putative protein in the compaction of hyphae in primordia.4 This protein was later purified and named aegerolysin.7 A detailed monitoring of the occurrence of ostreolysin and aegerolysin in mycelia and fruiting bodies of respective mushrooms (P. ostreatus and A. aegerita), and its correlation with the developmental stages of the fungi was performed using immunocytochemical methods. Both proteins were identified in the rapidly growing primordia, in the basidia and basidiospores of maturing fruiting bodies, which might indicate their function in initiation of fructification and/or sporulation.18 Their occurrence in peripheral parts of the fruiting bodies and lamellae suggests, in particular, that they may participate in differentiation of hyphae and formation of basidia and basidiospores. The latter hypothesis is supported by the finding that some bacterial aegerolysin-like proteins are expressed during sporulation.5 It is interesting to note that ostreolysin can markedly induce fructification of P. ostreatus even when applied externally to the mycelium.102

Perspectives

Although their physiological role in the producing organisms is not yet well understood, the biological properties of aegerolysin-like proteins make them interesting from several points of view. Asp-hemolysin, for example, is a lethal toxin that constitutes a very important part of the toxic battery of Aspergillus fumigatus during infection. This fungus causes a wide range of serious diseases, especially highly lethal invasive aspergillosis that often occurs in immunocompromised individuals, such as leukemia, cancer or AIDS patients, and after bone marrow and organ transplantations.103 Animal models have shown that anti-Asp-hemolysin IgG can successfully protect mice from the infection propagated with spores,98 and indicate that, beside the classical treatment with amphotericine, the use of anti-Asp-hemolysin IgG could be one of the strategies to protect immunocompromised patients from infection propagated by A. fumigatus. Moreover, the specific binding of Asp-hemolysin to ox-LDL makes this protein, its nonhemolytic recombinant mutants or synthetic forms86 a possible tool for investigation of the pathophysiological significance of ox-LDL.81 Further study on the binding mechanism between the Asp-hemolysin-related peptide and ox-LDL may provide important information on the prevention and treatment of atherosclerosis.

Specific binding to distinct membrane domains seems to be another characteristic feature of aegerolysin-like proteins, and has been most extensively studied in ostreolysin.51, 73, 74 Ostreolysin, and probably also other aegerolysins, specifically bind to raft-like cholesterol-rich membrane domains, and thus might be interesting as new tools for investigating these cell membrane domains. Their fluorescently- or spin-labeled mutants devoid of lytic activity could be useful in structural and functional studies of biological membranes. Indeed, several cytolytic proteins that specifically recognize particular components enriched in lipid rafts have been recently proposed as markers of raft-like membrane domains. A truncated, nontoxic mutant of lysenin (isolated from the earthworm Eisenia foetida) is a specific sphingomyelin-binding protein,104 cholera toxin B subunit (CT-B) binds to ganglioside G_M1,105 and a nontoxic derivative of perfringolysin O from Clostridium perfringens requires higher cholesterol content as a prerequisite for binding.106 In contrast to these cytolysins, ostreolysin was shown to specifically sense the combination of the two main lipid components of membrane rafts - cholesterol and sphingomyelin.51 Thus, this protein could be proposed as a useful external lipid raft marker, able to detect domains ascribed to cholesterol-sphingomyelin rafts. Ostreolysin does not colocalize with other raft-binding proteins like caveolin or the commercially available cholera-toxin B subunit.74 Hence, new insights on microheterogeneity of cholesterol-enriched membrane microdomains could be gained by using ostreolysin-like probes.

Last but not least, fungal aegerolysins are specifically expressed during the early phases of fructification.4, 7, 18 Stimulation of fruiting in mushrooms can be achieved by a variety of external stimuli, including substances of natural or synthetic origin, reduced temperature, light, and nutrient depletion.102 Better understanding of genes, pathways and cellular processes leading to the initiation and development of fruiting bodies in fungi may improve cultivation of edible mushrooms. In this regard, the ability of ostreolysin to enhance the fructification of oyster mushrooms, and possibly of other mushrooms when applied externally102 makes this protein and its genetically engineered recombinant forms interesting from the commercial and biotechnological points of view.

Supporting Information

References

1 Sakaguchi O, Shimada H Yokota K ( 1975) Purification and characteristics of hemolytic toxin from Aspergillus fumigatus. Jpn J Med Sci Biol 28: 328–331.
CAS PubMed Web of Science® Google Scholar
2 Ebina K, Sakagami H, Yokota K Kondo H ( 1994) Cloning and nucleotide sequence of cDNA encoding Asp-hemolysin from Aspergillus fumigatus. Biochim Biophys Acta 1219: 148–150.
10.1016/0167-4781(94)90258-5
CAS PubMed Web of Science® Google Scholar
3 Rementeria A, López-Molina N, Ludwig A, Vivanco AB, Bikandi J, Pontón J, Garaizar, J ( 2005) Genes and molecules involved in Aspergillus fumigatus virulence. Rev Iberoam Micol 22: 1–23.
10.1016/S1130-1406(05)70001-2
PubMed Google Scholar
4 Fernandez-Espinar MT, Labarère J ( 1997) Cloning and sequencing of the Aa-Pri1 gene specifically expressed during fruiting initiation in the edible mushroom Agrocybe aegerita, and analysis of the predicted amino-acid sequence. Curr Genet 32: 420–424.
10.1007/s002940050297
CAS PubMed Google Scholar
5 Barloy F, Lecadet MM, Delécluse A ( 1998) Cloning and sequencing of three new putative toxin genes from Clostridium bifermentans CH18. Gene 211: 293–299.
10.1016/S0378-1119(98)00122-X
CAS PubMed Web of Science® Google Scholar
6 Stover CK, Pham XQ, Erwin AL, Mizoguchi SD, Warrener P, Hickey MJ, Brinkman FS, Hufnagle WO, Kowalik DJ, Lagrou M, Garber RL, Goltry L, Tolentino E, Westbrock-Wadman S, Yuan Y, Brody LL, Coulter SN, Folger KR, Kas A, Larbig K, Lim R, Smith K, Spencer D, Wong GK, Wu Z, Paulsen IT, Reizer J, Saier MH, Hancock RE, Lory S Olson MV ( 2000) Complete genome sequence of Pseudomonas aeruginosa PA01, an opportunistic pathogen. Nature 406: 959–964.
10.1038/35023079
CAS PubMed Web of Science® Google Scholar
7 Berne S, Križaj I, Pohleven F, Turk T, Maček P, Sepčić K ( 2002) Pleurotus and Agrocybe hemolysins, new proteins hypothetically involved in fungal fruiting. Biochim Biophys Acta 1570: 153–159.
10.1016/S0304-4165(02)00190-3
CAS PubMed Web of Science® Google Scholar
8 Galagan JE, Calvo SE, Borkovich KA, Selker EU, Read ND, Jaffe D, FitzHugh W, Ma LJ, Smirnov S, Purcell S, Rehman B, Elkins T, Engels R, Wang S, Nielsen CB, Butler J, Endrizzi M, Qui D, Ianakiev P, Bell-Pedersen D, Nelson MA, Werner-Washburne M, Selitrennikoff CP, Kinsey JA, Braun EL, Zelter A, Schulte U, Kothe GO, Jedd G, Mewes W, Staben C, Marcotte E, Greenberg D, Roy A, Foley K, Naylor J, Stange-Thomann N, Barrett R, Gnerre S, Kamal M, Kamvysselis M, Mauceli E, Bielke C, Rudd S, Frishman D, Krystofova S, Rasmussen C, Metzenberg RL, Perkins DD, Kroken S, Cogoni C, Macino G, Catcheside D, Li W, Pratt RJ, Osmani SA, DeSouza CP, Glass L, Orbach MJ, Berglund JA, Voelker R, Yarden O, Plamann M, Seiler S, Dunlap J, Radford A, Aramayo R, Natvig DO, Alex LA, Mannhaupt G, Ebbole DJ, Freitag M, Paulsen I, Sachs MS, Lander ES, Nusbaum C, Birren B ( 2003) The genome sequence of the filamentous fungus. Neurospora crassa Nature 422: 859–868.
10.1038/nature01554
CAS PubMed Web of Science® Google Scholar
9 Tomita T, Noguchi K, Mimuro H, Ukaji F, Ito K, Sugawara-Tomita N, Hashimoto Y ( 2004) Pleurotolysin, a novel sphingomyelin-specific two-component cytolysin from the edible mushroom Pleurotus ostreatus, assembles into a transmembrane pore complex. J Biol Chem 279: 26975–26982.
10.1074/jbc.M402676200
CAS PubMed Web of Science® Google Scholar
10 Ngai PHK, Ng TB ( 2006) A hemolysin from the mushroom Pleurotus eryngii. Appl Microbiol Biotechnol 72: 1185–1191.
10.1007/s00253-006-0406-6
CAS PubMed Web of Science® Google Scholar
11 Rao J, DiGiandomenico A, Unger J, Bao Y, Polanowska-Grabowska RK, Goldberg JB ( 2008) A novel oxidized low-density lipoprotein-binding protein from Pseudomonas aeruginosa. Microbiology 154: 654–665.
10.1099/mic.0.2007/011429-0
CAS PubMed Web of Science® Google Scholar
12 Bodin E, Gautier L ( 1906) Note sur une toxine producte par l'Aspergillus fumigatus. Ann Inst Pasteur 20: 209–24.
Google Scholar
13 Henrici AT ( 1938) An endotoxin from Aspergillus fumigatus. J. Immunol 36: 319–338.
Google Scholar
14 Tilden EB, Hatton EH, Freeman S, Williamson WM, Koenig VL ( 1961) Preparation and properties of the endotoxins of Aspergillus fumigatus and Aspergillus flavus. Mycopathol Mycol Appl 14: 325–346.
10.1007/BF02051548
CAS PubMed Google Scholar
15 Tilden EB, Freeman S, Lombard L ( 1963) Further studies of the Aspergillus endotoxins. Mycopathol Mycol Appl 20: 253–271.
10.1007/BF02089213
CAS PubMed Google Scholar
16 Rau EM, Tilden EB, Koenig VL ( 1961) Partial purification and characterization of the endotoxin from Aspergillus fumigatus. Mycopathologia 14: 347–358.
10.1007/BF02051549
CAS PubMed Google Scholar
17 Bernheimer AW, Avigad LS ( 1979) Cytolytic protein from the edible mushroom, Pleurotus ostreatus. Biochim Biophys Acta 585: 451–461.
10.1016/0304-4165(79)90090-4
CAS PubMed Web of Science® Google Scholar
18 Vidic I, Berne S, Drobne D, Maček P, Frangež R, Turk T, Štrus J, Sepčić K ( 2005) Temporal and spatial expression of ostreolysin during development of the oyster mushroom (Pleurotus ostreatus). Mycol Res 109: 377–382.
10.1017/S0953756204002187
CAS PubMed Web of Science® Google Scholar
19 Kumagai T, Kudo Y, Fukuchi Y, Ebina K, Yokota K ( 2002) Expression of a synthetic gene encoding the Asp-hemolysin from Aspergillus fumigatus in Escherichia coli. Biol Pharm Bull 25: 115–117.
10.1248/bpb.25.115
CAS PubMed Web of Science® Google Scholar
20 Juárez-Pérez V, Delécluse A ( 2001) The Cry toxins and the putative hemolysins of Clostridium bifermentans ser. malaysia are not involved in mosquitocidal activity. J Invertebr Pathol 78: 57–58.
10.1006/jipa.2001.5042
CAS PubMed Web of Science® Google Scholar
21 Sakurai N, Kaneko J, Kamio Y, Tomita T ( 2004) Cloning, expression, and pore-forming properties of mature and precursor forms of pleurotolysin, a sphingomyelin-specific two-component cytolysin from the edible mushroom Pleurotus ostreatus. Biochim Biophys Acta 1679: 65–73.
10.1016/j.bbaexp.2004.05.002
CAS PubMed Web of Science® Google Scholar
22 Finn RD, Tate J, Mistry J, Coggill PC, Sammut JS, Hotz HR, Ceric G, Forslund K, Eddy SR, Sonnhammer EL, Bateman A ( 2008) The Pfam protein families database. Nucleic Acids Res 36(Database Issue): D281–D288.
10.1093/nar/gkm960
CAS PubMed Web of Science® Google Scholar
23 Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W Lipman DJ ( 1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25: 3389–3402.
10.1093/nar/25.17.3389
CAS PubMed Web of Science® Google Scholar
24 Schäffer AA, Aravind L, Madden TL, Shavirin S, Spouge JL, Wolf YI, Koonin EV, Altschul SF ( 2001) Improving the accuracy of PSI-BLAST protein database searches with composition-based statistics and other refinements. Nucleic Acids Res 29: 2994–3005.
10.1093/nar/29.14.2994
CAS PubMed Web of Science® Google Scholar
25 Wallace IM, O'Sullivan O, Higgins DG, Notredame C ( 2006) M-Coffee: combining multiple sequence alignment methods with T-Coffee. Nucleic Acids Res 34: 1692–1699.
10.1093/nar/gkl091
CAS PubMed Web of Science® Google Scholar
26 Moretti S, Armougom F, Wallace IM, Higgins DG, Jongeneel CV, Notredame C ( 2007) The M-Coffee web server: a meta-method for computing multiple sequence alignments by combining alternative alignment methods. Nucleic Acids Res 35 (Web Server issue): W645–W648.
10.1093/nar/gkm333
PubMed Web of Science® Google Scholar
27 Nicholas KB, Nicholas HB,Jr, Deerfield DWII ( 1997) GeneDoc: analysis and visualization of genetic variation, EMBNEW.NEWS 4: 14. Available at: http://www.psc.edu/biomed/genedoc.
Google Scholar
28 Clamp M, Cuff J, Searle SM, Barton GJ ( 2004) The jalview java alignment editor. Bioinformatics 20: 426–427.
10.1093/bioinformatics/btg430
CAS PubMed Web of Science® Google Scholar
29 Cole C, Barber JD, Barton GJ ( 2008) The Jpred 3 secondary structure prediction server. Nucleic Acids Res 36 (Web Server issue): W197–W201.
10.1093/nar/gkn238
CAS PubMed Web of Science® Google Scholar
30 Lin H-N, Chang J-M, Wu K-P, Sung T-Y, Hsu W-L ( 2005) HYPROSP II-A knowledge-based hybrid method for protein secondary structure prediction based on local prediction confidence. Bioinformatics 21: 3227–3233.
10.1093/bioinformatics/bti524
CAS PubMed Web of Science® Google Scholar
31 Rost B, Yachdav G, Liu J ( 2004) The PredictProtein server. Nucleic Acids Res 32 (Web Server issue): W321–W326.
10.1093/nar/gkh377
CAS PubMed Web of Science® Google Scholar
32 Berne S, Sepčić K, Anderluh G, Turk T, Maček P, Poklar Ulrih N ( 2005) Effect of pH on the pore forming activity and conformational stability of ostreolysin, a lipid raft-binding protein from the edible mushroom Pleurotus ostreatus. Biochemistry 44: 11137–11147.
10.1021/bi051013y
CAS PubMed Web of Science® Google Scholar
33 Sepčić K, Berne S, Potrich C, Turk T, Maček P, Menestrina G ( 2003) Interaction of ostreolysin, a cytolytic protein from the edible mushroom Pleurotus ostreatus, with lipid membranes and modulation by lysophospholipids. Eur J Biochem 270: 1199–1210.
10.1046/j.1432-1033.2003.03480.x
CAS PubMed Web of Science® Google Scholar
34 Fukuchi Y, Kumagai T, Ebina K, Yokota K ( 1996b) Apolipoprotein B inhibits the hemolytic activity of asp-hemolysin from Aspergillus fumigatus. Biol Pharm Bull 19: 547–550.
10.1248/bpb.19.547
CAS PubMed Web of Science® Google Scholar
35 Rice P ( 2000) Longden I Bleasby A. EMBOSS: The european molecular biology open software suite. Trends Genet 16: 276–277.
10.1016/S0168-9525(00)02024-2
CAS PubMed Web of Science® Google Scholar
36 Fukuchi Y, Kudo Y, Kumagai T, Ebina K, Yokota K ( 1996a) Binding assay of low density lipoprotein to Asp-hemolysin from Aspergillus fumigatus. Biol Pharm Bull 19: 1380–1381.
10.1248/bpb.19.1380
CAS PubMed Web of Science® Google Scholar
37 Fukuchi Y, Kudo Y, Kumagai T, Ebina K, Yokota K ( 1998) Oxidized low density lipoprotein inhibits the hemolytic activity of Asp-hemolysin from Aspergillus fumigatus. FEMS Microbiol Lett 167: 275–280.
10.1111/j.1574-6968.1998.tb13239.x
CAS PubMed Web of Science® Google Scholar
38 Kudo Y, Ootani T, Kumagai T, Fukuchi Y, Ebina K ( 2002) A novel oxidized low-density lipoprotein binding protein, Asp-hemolysin, recognizes lysophosphatidylcholine. Biol Pharm Bull 25: 787–790.
10.1248/bpb.25.787
CAS PubMed Web of Science® Google Scholar
39 Kamaguchi A, Yokota K, Sakaguchi O ( 1979) Investigation of the hemolytic site of Asp-hemolysin. Jpn J Med Sci Biol 32: 118–121.
CAS PubMed Web of Science® Google Scholar
40 Hong Q, Gutiérrez-Aguirre I, Barlič A, Malovrh P, Kristan K, Podlesek Z, Maček P, Turk D, González-Mañas JH, Lakey JH, Anderluh G ( 2002) Two-step membrane binding by equinatoxin II, a pore-forming toxin from the sea anemone, involves an exposed aromatic cluster and a flexible helix. J Biol Chem 277: 41916–41924.
10.1074/jbc.M204625200
CAS PubMed Web of Science® Google Scholar
41 Ramachandran R, Heuck AP, Tweten RK, Johnson AE ( 2002) Structural insights into the membrane-anchoring mechanism of a cholesterol-dependent cytolysin. Nat Struct Biol 9: 823–827.
CAS PubMed Web of Science® Google Scholar
42 Palmer M ( 2004) Cholesterol and the activity of bacterial toxins. FEMS Microbiol Lett 238: 281–289.
10.1111/j.1574-6968.2004.tb09768.x
CAS PubMed Web of Science® Google Scholar
43 Hedges SB ( 2002) The origin and evolution of model organisms. Nature Rev Gen 3: 838–849.
10.1038/nrg929
CAS PubMed Web of Science® Google Scholar
44 Abby S, Daubin V ( 2007) Comparative genomics and the evolution of prokaryotes. Trends Microbiol 15: 135–141.
10.1016/j.tim.2007.01.007
CAS PubMed Web of Science® Google Scholar
45 Keeling PJ, Burger G, Durnford DG, Lang BF, Lee RW, Pearlman RE, Roger AJ, Gray MW ( 2005) The tree of eukaryotes. Trends Ecol Evol 20: 670–676.
10.1016/j.tree.2005.09.005
PubMed Web of Science® Google Scholar
46 Sher D, Fishman Y, Zhang M, Lebendiker M, Gaathon A, Mancheño JM, Zlotkin E ( 2005) Hydralysins, a new category of beta-pore-forming toxins in cnidaria. J Biol Chem 80: 22847–22855.
10.1074/jbc.M503242200
CAS Web of Science® Google Scholar
47 Cho YS, Kim JS, Crowley DE, Cho BG ( 2003) Growth promotion of the edible fungus Pleurotus ostreatus by fluorescent pseudomonads. FEMS Microbiol Lett 218: 271–276.
10.1016/S0378-1097(02)01144-8
CAS PubMed Web of Science® Google Scholar
48 Richardson AO, Palmer JD ( 2007) Horizontal gene transfer in plants. J Exp Bot 58: 1–9.
10.1093/jxb/erl148
CAS PubMed Web of Science® Google Scholar
49 Richards TA, Dacks JB, Jenkinson JM, Thornton CR, Talbot NJ ( 2006) Evolution of filamentous plant pathogens: gene exchange across eukaryotic kingdoms. Curr Biol 16: 1857–1864.
10.1016/j.cub.2006.07.052
CAS PubMed Web of Science® Google Scholar
50 Berne S ( 2004) Biochemical and cytolytic properties of a protein from oyster mushroom Pleurotus ostreatus (Jacq. ex. Fr.) Kumm. Ph. D. thesis, Biotechnical faculty, Ljubljana.
Google Scholar
51 Karlsson M, Olson A, Stenlid J ( 2003) Expressed sequences from the basidiomycetous tree pathogen Heterobasidion annosum during early infection of Scots pine. Fungal Genet Biol 39: 51–59.
10.1016/S1087-1845(02)00586-8
CAS PubMed Web of Science® Google Scholar
52 Sepčić K, Berne S, Rebolj K, Batista U, Plemenitaš A, Šentjurc M, Maček P ( 2004) Ostreolysin, a pore-forming protein from the oyster mushroom, interacts specifically with membrane cholesterol-rich lipid domains. FEBS Lett 575: 81–85.
10.1016/j.febslet.2004.07.093
CAS PubMed Web of Science® Google Scholar
53 Nierman WC, Pain A, Anderson MJ, Wortman JR, Kim HS, Arroyo J, Berriman M, Abe K, Archer DB, Bermejo C, Bennett J, Bowyer P, Chen D, Collins M, Coulsen R, Davies R, Dyer PS, Farman M, Fedorova N, Fedorova N, Feldblyum TV, Fischer R, Fosker N, Fraser A, Garcia JL, Garcia MJ, Goble A, Goldman GH, Gomi K, Griffith-Jones S, Gwilliam R, Haas B, Haas H, Harris D, Horiuchi H, Huang J, Humphray S, Jimenez J, Keller N, Khouri H, Kitamoto K, Kobayashi T, Konzack S, Kulkarni R, Kumagai T, Lafton A, Latgé JP, Li W, Lord A, Lu C, Majoros WH, May GS, Miller BL, Mohamoud Y, Molina M, Monod M, Mouyna I, Mulligan S, Murphy L, O'Neil S, Paulsen I, Penalva MA, Pertea M, Price C, Pritchard BL, Quail MA, Rabbinowitsch E, Rawlins N, Rajandream MA, Reichard U, Renauld H, Robson GD, Rodriguez de Cordoba S, Rodriguez-Pena JM, Ronning CM, Rutter S, Salzberg SL, Sanchez M, Sanchez-Ferrero JC, Saunders D, Seeger K, Squares R, Squares S, Takeuchi M, Tekaia F, Turner G, Vazquez de Aldana CR, Weidman J, White O, Woodward J, Yu JH, Fraser C, Galagan JE, Asai K, Machida M, Hall N, Barrell B, Denning DW ( 2005) Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus. Nature 438: 1151–1156.
10.1038/nature04332
CAS PubMed Web of Science® Google Scholar
54 Pel HJ, de Winde JH, Archer DB, Dyer PS, Hofmann G, Schaap PJ, Turner G, de Vries RP, Albang R, Albermann K, Andersen MR, Bendtsen JD, Benen JA, van den Berg M, Breestraat S, Caddick MX, Contreras R, Cornell M, Coutinho PM, Danchin EG, Debets AJ, Dekker P, van Dijck PW, van Dijk A, Dijkhuizen L, Driessen AJ, d'Enfert C, Geysens S, Goosen Groot GS, de Groot PW, Guillemette T, Henrissat B, Herweijer M, van den Hombergh JP, van den Hondel CA, van der Heijden RT, van der Kaaij RM, Klis FM, Kools HJ, Kubicek CP, van Kuyk PA, Lauber J, Lu X, van der Maarel MJ, Meulenberg R, Menke H, Mortimer MA, Nielsen J, Oliver SG, Olsthoorn M, Pal K, van Peij NN, Ram AF, Rinas U, Roubos JA, Sagt CM, Schmoll M, Sun J, Ussery D, Varga J, Vervecken W, van de Vondervoort PJ, Wedler H, Wosten HA, Zeng AP, van Ooyen AJ, Visser J Stam H ( 2007) Genome sequencing and analysis of the versatile cell factory Aspergillus niger CBS 513.88. Nat Biotechnol 25: 221–231.
10.1038/nbt1282
PubMed Web of Science® Google Scholar
55 Machida M, Asai K, Sano M, Tanaka T, Kumagai T, Terai G, Kusumoto K, Arima T, Akita O, Kashiwagi Y, Abe K, Gomi K, Horiuchi H, Kitamoto K, Kobayashi T, Takeuchi M, Denning DW, Galagan JE, Nierman WC, Yu J, Archer DB, Bennett JW, Bhatnagar D, Cleveland TE, Fedorova ND, Gotoh O, Horikawa H, Hosoyama A, Ichinomiya M, Igarashi R, Iwashita K, Juvvadi PR, Kato M, Kato Y, Kin T, Kokubun A, Maeda H, Maeyama N, Maruyama J, Nagasaki H, Nakajima T, Oda K, Okada K, Paulsen I, Sakamoto K, Sawano T, Takahashi M, Takase K, Terabayashi Y, Wortman JR, Yamada O, Yamagata Y, Anazawa H, Hata Y, Koide Y, Komori T, Koyama Y, Minetoki T, Suharnan S, Tanaka A, Isono K, Kuhara S, Ogasawara N, Kikuchi H ( 2005) Genome sequencing and analysis of Aspergillus oryzae. Nature 438: 1157–1161.
10.1038/nature04300
PubMed Web of Science® Google Scholar
56 Silva EM, Valdes J, Holmes D, Shmaryahu A, Valenzuela PD ( 2006) Generation and analysis of expressed sequence tags from Botrytis cinerea. Biol Res 39: 367–379.
10.4067/S0716-97602006000200018
CAS PubMed Web of Science® Google Scholar
57 Felipe MS, Andrade RV, Arraes FB, Nicola AM, Maranhao AQ, Torres FA, Silva-Pereira I, Pocas-Fonseca MJ, Campos EG, Moraes LM, Andrade PA, Tavares AH, Silva SS, Kyaw CM, Souza DP, Network P, Pereira M, Jesuino RS, Andrade EV, Parente JA, Oliveira GS, Barbosa MS, Martins NF, Fachin AL, Cardoso RS, Passos GA, Almeida NF, Walter ME, Soares CM, Carvalho MJ Brigido MM ( 2005) Transcriptional Profiles of the Human Pathogenic Fungus Paracoccidioides brasiliensis in Mycelium and Yeast Cells. J Biol Chem 280: 24706–24714.
10.1074/jbc.M500625200
PubMed Web of Science® Google Scholar
58 Herwig R, Schulz B, Weisshaar B, Hennig S, Steinfath M, Drungowski M, Stahl D, Wruck W, Menze A, O'Brien J, Lehrach H, Radelof U ( 2002) Construction of a ‘unigene’ cDNA clone set by oligonucleotide fingerprinting allows access to 25 000 potential sugar beet genes. Plant J 32: 845–857.
10.1046/j.1365-313X.2002.01457.x
CAS PubMed Web of Science® Google Scholar
59 Baum JA, Chu CR, Rupar M, Brown GR, Donovan WP, Huesing JE, Ilagan O, Malvar TM, Pleau M, Walters M, Vaughn T ( 2004) Binary toxins from Bacillus thuringiensis active against the western corn rootworm, Diabrotica virgifera virgifera LeConte. Appl Environ Microbiol 70: 4889–4898.
10.1128/AEM.70.8.4889-4898.2004
CAS PubMed Web of Science® Google Scholar
60 Schnepf HE, Lee S, Dojillo J, Burmeister P, Fencil K, Morera L, Nygaard L, Narva KE, Wolt JD ( 2005) Characterization of Cry34/Cry35 binary insecticidal proteins from diverse Bacillus thuringiensis strain collections. Appl Environ Microbiol 71: 1765–1774.
10.1128/AEM.71.4.1765-1774.2005
CAS PubMed Web of Science® Google Scholar
61 Wang L, Xue J, Seaborn CP, Arif BM, Cheng XW ( 2006) Sequence and organization of the Trichoplusia ni ascovirus 2c (Ascoviridae) genome. Virology 354: 167–177.
10.1016/j.virol.2006.06.029
CAS PubMed Web of Science® Google Scholar
62 Ebina K, Ichinowatari S, Yokota K ( 1985) Studies on toxin of Aspergillus fumigatus. XXII Fashion of binding of Asp-hemolysin to human erythrocytes and Asp-hemolysin-binding proteins of erythrocyte membranes Microbiol Immunol 29: 91–101.
CAS PubMed Web of Science® Google Scholar
63 Yokota K, Kamaguchi A, Sakaguchi O ( 1984a) Studies on the toxin of Aspergillus fumigatus XVIII. Photooxidation of asp-hemolysin in the presence of various dyes and its relation to the site of hemolytic activity.Microbiol Immunol 28: 385–391.
CAS PubMed Web of Science® Google Scholar
64 Yokota K, Ichinowatari S, Ebina K ( 1984b) Studies on the toxin of Aspergillus fumigatus XX. Chemical modification of Asp-hemolysin. Jpn J Med Mycol 25: 332–339.
10.3314/jjmm1960.25.332
CAS Google Scholar
65 Yokota K, Ichinowatari S, Ebina K, Wakabayashi N ( 1985) Studies on the toxin of Aspergillus fumigatus XXI. Site of binding of Asp-hemolysin to erythrocytes and mechanism of inhibition of hemolysis. Jpn J Med Mycol 26: 332–339.
10.3314/jjmm1960.26.70
Google Scholar
66 Sakaguchi O, Yokota K, Kamaguchi A ( 1977) Studies on the toxin of Aspergillus fumigatus VIII. Biological properties of Asp-hemolysin. Nippon Saikingaku Zasshi 32: 821–828.
10.3412/jsb.32.821
CAS PubMed Google Scholar
67 Rebolj K, Sepčić K ( 2008) Ostreolysin, a cytolytic protein from culinary-medicinal oyster mushroom Pleurotus ostreatus (Jack.: Fr.) P. Kumm. (Agaricomycetidae), and its potential use in medicine and biotechnology. Int J Med Mushr 10: 293–302.
10.1615/IntJMedMushr.v10.i4.10
CAS Web of Science® Google Scholar
68 Žužek MC, Maček P, Sepčić K, Cestnik V, Frangež R ( 2006) Toxic and lethal effects of ostreolysin, a cytolytic protein from edible oyster mushroom (Pleurotus ostreatus), in rodents. Toxicon 48: 264–271.
10.1016/j.toxicon.2006.05.011
CAS PubMed Web of Science® Google Scholar
69 Simons K, Ikonen E. Functional rafts in cell membranes. Nature 387: 569–572.
Google Scholar
70 Edidin M ( 2003) The state of lipid rafts: from model membranes to cells. Annu Rev Biophys Biomol Struct 32: 257–283.
10.1146/annurev.biophys.32.110601.142439
CAS PubMed Web of Science® Google Scholar
71 London E ( 2002) Insights into lipid raft structure and formation from experiments in model membranes. Curr Opin Struct Biol 12: 480–486.
10.1016/S0959-440X(02)00351-2
CAS PubMed Web of Science® Google Scholar
72 Lichtenberg D, Goni FM, Heerklotz H ( 2005) Detergent-resistant membranes should not be identified with membrane rafts. Trends Biochem Sci 30: 430–436.
10.1016/j.tibs.2005.06.004
CAS PubMed Web of Science® Google Scholar
73 de Almeida RFM, Fedorov A, Prieto M ( 2003) Sphingomyelin/phosphatidylcholine/cholesterol phase diagram: boundaries and composition of lipid rafts. Biophys J 85: 2406–2416.
10.1016/S0006-3495(03)74664-5
CAS PubMed Web of Science® Google Scholar
74 Rebolj K, Poklar Ulrih N, Maček P, Sepčić K ( 2006) Steroid structural requirements for interaction of ostreolysin, a lipid-raft binding cytolysin, with lipid monolayers and bilayers. Biochim Biophys Acta 1758: 1662–1670.
10.1016/j.bbamem.2006.06.003
CAS PubMed Web of Science® Google Scholar
75 Chowdhury HH, Rebolj K, Kreft M, Zorec R, Maček P, Sepčić K ( 2008) Lysophospholipids prevent binding of a cytolytic protein ostreolysin to cholesterol-enriched membrane domains. Toxicon 51: 1345–1356.
10.1016/j.toxicon.2008.03.010
CAS PubMed Web of Science® Google Scholar
76 Hammond AT, Heberle FA, Baumgart T, Holowka D, Baird B, Feigenson GW ( 2005) Crosslinking a lipid raft component triggers liquid ordered-liquid disordered phase separation in model plasma membranes. Proc Natl Acad Sci USA 102: 6320–3625.
10.1073/pnas.0405654102
CAS PubMed Web of Science® Google Scholar
77 Nagamune H, Ohnishi C, Katsuura A, Fushitani K, Whiley RA, Tsuji A, Matsuda Y ( 1996) Intermedilysin, a novel cytotoxin specific for human cells secreted by Streptococcus intermedius UNS46 isolated from a human liver abscess. Infect Immun 64: 3093–3100.
10.1128/IAI.64.8.3093-3100.1996
CAS PubMed Web of Science® Google Scholar
78 Giddings KS, Johnson AE, Tweten R ( 2003) Redefining cholesterol's role in the mechanism of the cholesterol-dependent cytolysins. Proc Natl Acad Sci USA 100: 11315–11320.
10.1073/pnas.2033520100
CAS PubMed Web of Science® Google Scholar
79 Polekhina G, Giddings KS, Tweten RK, Parker MW ( 2005) Insights into the action of the superfamily of cholesterol-dependent cytolysins from studies of intermedilysin. Proc Natl Acad Sci USA 102: 600–605.
10.1073/pnas.0403229101
CAS PubMed Web of Science® Google Scholar
80 Tweten R ( 2005) Cholesterol-dependent cytolysins, a family of versatile pore-forming toxins. Infect Immun 73: 6199–6209.
10.1128/IAI.73.10.6199-6209.2005
CAS PubMed Web of Science® Google Scholar
81 Olson R, Gouaux E ( 2005) Crystal structure of the Vibrio cholerae cytolysin (VCC) pro-toxin and its assembly into a heptameric transmembrane pore. J Mol Biol 350: 997–1016.
10.1016/j.jmb.2005.05.045
CAS PubMed Web of Science® Google Scholar
82 Zitzer A, Zitzer O, Bhakdi S, Palmer M ( 1999) Oligomerization of Vibrio cholerae cytolysin yields a pentameric pore and has a dual specificity for cholesterol and sphingolipids in the target membrane. J Biol Chem 274: 1375–1380.
10.1074/jbc.274.3.1375
CAS PubMed Web of Science® Google Scholar
83 Kudo Y, Fukuchi Y, Kumagai T, Ebina K Yokota K ( 2001) Oxidized low-density lipoprotein-binding specificity of Asp-hemolysin from Aspergillus fumigatus. Biochim Biophys Acta 1568: 183–188.
10.1016/S0304-4165(01)00217-3
CAS PubMed Web of Science® Google Scholar
84 Kumagai T, Nagata Y, Kudo Y, Fukuchi Y, Ebina K Yokota K ( 1999) Cytotoxic activity and cytokine gene induction of Asp-hemolysin to murine macrophages. Jpn J Med Mycol 40: 217–222.
10.3314/jjmm.40.217
CAS PubMed Google Scholar
85 Fukuchi Y ( 2001) Interactions between Asp-hemolysin from Aspergillus fumigatus and blood plasma components. Yakugaku Zasshi 121: 423–432.
10.1248/yakushi.121.423
CAS PubMed Web of Science® Google Scholar
86 Kudo Y, Kumagai T, Fukuchi Y, Ebina K, Yokota K ( 1999) Binding of Asp-hemolysin from Aspergillus fumigatus to oxidized low density lipoprotein. Biol Pharm Bull 22: 549–550.
10.1248/bpb.22.549
CAS PubMed Web of Science® Google Scholar
87 Kumagai T, Ogawa N, Tsutsumi H, Ebina K Yokota K ( 2005) A synthetic peptide (P-21) derived from Asp-hemolysin inhibits the induction of macrophage proliferation by oxidized low-density lipoprotein. Biol Pharm Bull 28: 1381–1384.
10.1248/bpb.28.1381
CAS PubMed Web of Science® Google Scholar
88 Kumagai T, Tsutsumi H, Ogawa N, Naito S, Ebina, K. Yokota K Nagata K ( 2006) Oxidized low-density lipoprotein-binding specificity of the Asp-hemolysin-related synthetic peptides from Aspergillus fumigatus. Biol Pharm Bull 29: 2181–2186.
10.1248/bpb.29.2181
CAS PubMed Web of Science® Google Scholar
89 Sakai M, Miyazaki A, Hakamata H, Sasaki T, Yui S, Yamazaki M, Shichiri M, Horiuchi S ( 1994) Lysophosphatidylcholine plays an essential role in the mitogenic effect of oxidized low density lipoprotein on murine macrophages. J Biol Chem 269: 31430–31435.
10.1016/S0021-9258(18)31712-5
CAS PubMed Web of Science® Google Scholar
90 Ebina K, Yokota K, Sakaguchi O ( 1983) Studies on toxin of Aspergilus fumigatus XVI. Biological properties of Asp-hemolysin as a parasite factor. Jpn J Med Mycol 24: 245–252.
10.3314/jjmm1960.24.245
CAS Google Scholar
91 Kumagai T, Nagata T, Kudo Y, Fukuchi Y, Ebina K Yokota K ( 2001) Cytotoxic activity and cytokine induction of Asp-hemolysin to vascular endothelial cells. Yakugaku Zasshi 121: 271–275.
10.1248/yakushi.121.271
CAS PubMed Web of Science® Google Scholar
92 Maličev E, Chowdhury H, Maček P, Sepčić K ( 2007) Effect of ostreolysin, an Asp-hemolysin isoform, on human chondrocytes and osteoblasts, and possible role of Asp-hemolysin in pathogenesis. Med Mycol 45: 123–30.
10.1080/13693780601039615
CAS PubMed Web of Science® Google Scholar
93 Rebolj K, Batista U, Sepčić K, Cestnik V, Maček P, Frangež R ( 2007) Ostreolysin affects rat aorta ring tension and endothelial cells viability in vitro. Toxicon 49: 1211–1213.
10.1016/j.toxicon.2007.01.016
CAS PubMed Web of Science® Google Scholar
94 Henrici AT ( 1938) An endotoxin from Aspergillus fumigatus. J Immunol 36: 319–338.
Google Scholar
95 Tilden EB, Hatton EH, Freeman S, Williamson WM, Koenig VL ( 1961) Preparation and properties of the endotoxins of Aspergillus fumigatus and Aspergillus flavus. Mycopathol. Mycol Appl 14: 325–346.
10.1007/BF02051548
CAS PubMed Google Scholar
96 Sakaguchi O, Yokota K ( 1972) Studies on the toxin of Aspergillus fumigatus III. Fraction causing dermonecrosis and hemorrhage-like changes. Nippon Saikingaku Zasshi 27: 649–655.
10.3412/jsb.27.649
CAS PubMed Google Scholar
97 Tilden EB, Freeman S, Lombard L ( 1963) Further studies of the Aspergillus endotoxins. Mycopathol. Mycol. Appl. 20: 253–271.
10.1007/BF02089213
CAS PubMed Google Scholar
98 Budzko DB, Negroni R ( 1976) Depletion of complemeni in vivo and in vitro by extracts of Aspergillus fumigatus. Int Arch Allergy Appl Immun 51: 518–524.
10.1159/000231625
CAS PubMed Web of Science® Google Scholar
99 Ebina K, Yokota K, Sakaguchi O ( 1982) Studies on toxin of Aspergillus fumigatus.. XIV Relationship between Asp-hemolysin and experimental infection for mice. Jpn J Med Mycol 23: 246–252.
10.3314/jjmm1960.23.246
Google Scholar
100 Ebina K, Ichinowatari S, Yokota K, Sakaguchi O ( 1984) Studies on toxin of Aspergilus fumigatus XIX. Biochemical alteration of sera after Asp-hemolysin inoculation or Aspergillus infection in mice. Jpn J Med Mycol 25: 236–243.
10.3314/jjmm1960.25.236
CAS Google Scholar
101 Al-Deen IHS, Twaij HAA, Al-Badr AA, Istarabad TAW ( 1987) Toxicologic and histopathologic studies of Pleurotus ostreatus mushroom in mice. J Ethnopharm 21: 297–305.
10.1016/0378-8741(87)90105-X
CAS PubMed Web of Science® Google Scholar
102 Schachter EN, Zuskin E, Goswami S, Castranova V, Arumugam U, Whitmer M, Siegel P, Chiarelli A Fainberg J ( 2005) Pharmacological study of oyster mushroom (Pleurotus ostreatus) extract on isolated guinea pig trachea smooth muscle. Lung 183: 63–71.
10.1007/s00408-004-2527-y
CAS PubMed Web of Science® Google Scholar
103 Berne S, Pohleven J, Vidic I, Rebolj K, Pohleven F, Turk T, Maček P, Sonnenberg A, Sepčić K ( 2007) Ostreolysin enhances fruiting initiation in the oyster mushroom (Pleurotus ostreatus). Mycol Res 111: 1431–1436.
10.1016/j.mycres.2007.09.005
CAS PubMed Web of Science® Google Scholar
104 Latgé JP ( 1999) Aspergillus fumigatus and aspergillosis. Clin Microbiol Rev 12: 310–350.
10.1128/CMR.12.2.310
CAS PubMed Web of Science® Google Scholar
105 Hullin-Matsuda F, Kobayashi T ( 2007) Monitoring the distribution and dynamics of signaling microdomains in living cells with lipid-specific probes. Cell Mol Life Sci 64: 2492–2504.
10.1007/s00018-007-7281-x
CAS PubMed Web of Science® Google Scholar
106 Chinnapen DJ, Chinnapen H, Saslowsky D Lencer WI ( 2007) Rafting with cholera toxin: endocytosis and trafficking from plasma membrane to ER. FEMS Microbiol Lett 266: 129–137.
10.1111/j.1574-6968.2006.00545.x
CAS PubMed Web of Science® Google Scholar
107 Nelson LD, Johnson AE, London E ( 2008) How interaction of perfringolysin O with membranes is controlled by sterol structure, lipid structure, and physiological low pH. J Biol Chem 283: 4632–4642.
10.1074/jbc.M709483200
CAS PubMed Web of Science® Google Scholar
108 Galagan JE, Calvo SE, Cuomo C, Ma LJ, Wortman JR, Batzoglou S, Lee SI, Basturkmen M, Spevak CC, Clutterbuck J, Kapitonov V, Jurka J, Scazzocchio C, Farman M, Butler J, Purcell S, Harris S, Braus GH, Draht O, Busch S, D'Enfert C, Bouchier C, Goldman GH, Bell-Pedersen D, Griffiths-Jones S, Doonan JH, Yu J, Vienken K, Pain A, Freitag M, Selker EU, Archer DB, Penalva MA, Oakley BR, Momany M, Tanaka T, Kumagai T, Asai K, Machida M, Nierman WC, Denning DW, Caddick M, Hynes M, Paoletti M, Fischer R, Miller B, Dyer P, Sachs MS, Osmani SA, Birren BW ( 2005) Sequencing of Aspergillus nidulans and comparative analysis with A. fumigatus and A. oryzae. Nature 438: 1105–1115.
10.1038/nature04341
CAS PubMed Web of Science® Google Scholar
109 Lee SH, Kim BG, Kim KJ, Lee JS, Yun DW, Hahn JH, Kim GH, Lee KH, Suh DS, Kwon ST, Lee CS Yoo YB ( 2002) Comparative analysis of sequences expressed during the liquid-cultured mycelia and fruit body stages of Pleurotus ostreatus. Fungal Genet Biol 35: 115–134.
10.1006/fgbi.2001.1310
CAS PubMed Web of Science® Google Scholar

Citing Literature

Volume18, Issue4

April 2009

Pages 694-706

Aegerolysins: Structure, function, and putative biological role

Abstract

Introduction

Purification of Native and Recombinant Aegerolysin-Like Proteins