Near-Stoichiometric interaction between the non-specific lipid-transfer protein of the yeast Candida tropicalis and peroxisomal acyl-coenzyme A oxidase prevents the thermal denaturation of the enzyme in vitro
Toshiro Niki
Faculty of Integrated Arts and Sciences, Hiroshima University, Kagamiyama, Higashi-Hiroshima 724, Japan
Search for more papers by this authorMasanori Bun-Ya
Faculty of Integrated Arts and Sciences, Hiroshima University, Kagamiyama, Higashi-Hiroshima 724, Japan
Search for more papers by this authorYoshitaka Muro
Faculty of Integrated Arts and Sciences, Hiroshima University, Kagamiyama, Higashi-Hiroshima 724, Japan
Search for more papers by this authorCorresponding Author
Tatsuyuki Kamiryo
Faculty of Integrated Arts and Sciences, Hiroshima University, Kagamiyama, Higashi-Hiroshima 724, Japan
Faculty of Integrated Arts and Sciences, Hiroshima University, Kagamiyama, Higashi-Hiroshima 724, JapanSearch for more papers by this authorYoshikazu Hiraga
Faculty of Science, Hiroshima University, Kagamiyama, Higashi-Hiroshima 724, Japan
Search for more papers by this authorToshiro Niki
Faculty of Integrated Arts and Sciences, Hiroshima University, Kagamiyama, Higashi-Hiroshima 724, Japan
Search for more papers by this authorMasanori Bun-Ya
Faculty of Integrated Arts and Sciences, Hiroshima University, Kagamiyama, Higashi-Hiroshima 724, Japan
Search for more papers by this authorYoshitaka Muro
Faculty of Integrated Arts and Sciences, Hiroshima University, Kagamiyama, Higashi-Hiroshima 724, Japan
Search for more papers by this authorCorresponding Author
Tatsuyuki Kamiryo
Faculty of Integrated Arts and Sciences, Hiroshima University, Kagamiyama, Higashi-Hiroshima 724, Japan
Faculty of Integrated Arts and Sciences, Hiroshima University, Kagamiyama, Higashi-Hiroshima 724, JapanSearch for more papers by this authorYoshikazu Hiraga
Faculty of Science, Hiroshima University, Kagamiyama, Higashi-Hiroshima 724, Japan
Search for more papers by this authorAbstract
A 14-kDa peroxisomal-matrix protein, named PXP-18, of the yeast Candida tropicalis is a structural and functional homologue of the mammalian nonspecific lipid-transfer protein (identical to sterol carrier protein-2). PXP-18 protected acyl-coenzyme A oxidase (ACO), the rate limiting enzyme of the peroxisomal β-oxidation of fatty acids, from thermal inactivation at 48°C or 70°C. This effect was dose-dependent and not replaceable either by chicken egg white lysozyme, which is similar to PXP-18 (insofar as it is basic, small, and monomeric), or by bovine serum albumin, a carrier of lipids in the blood. ACO was irreversibly denatured by heat treatment at 70°C for 15 min. However, when ACO and PXP-18 were similarly heat-treated, they formed a large complex at a molar ratio of PXP-18 to ACO subunit that was about one, independent of their initial ratio. This near-stoichiometric complex had ACO activity after a 500-fold dilution and was accompanied by ACO that was free of PXP-18 and indistinguishable from native ACO in size and activity. PXP-18 also protected urate oxidase, another peroxisomal enzyme, from inactivation at 66°C for 15 min and facilitated the renaturation of ACO denatured by 2 M urea. These results indicated that PXP-18 is active in modulating the structure of peroxisomal enzymes in vitro. It is possible that PXP-18 functions as a stress protein or as a part of the system that keeps peroxisomal proteins intact.
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