A novel recombinant dual human SCF expressed in and purified from silkworm, Bombyx mori, possesses higher bioactivity than recombinant monomeric human SCF
Junhai Han
School of Life Science and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China
Search for more papers by this authorYuhui Zang
School of Life Science and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China
Search for more papers by this authorHaiqin Lu
School of Life Science and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China
Search for more papers by this authorJie Zhu
School of Life Science and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China
Search for more papers by this authorJunchuan Qin
School of Life Science and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China
Search for more papers by this authorJunhai Han
School of Life Science and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China
Search for more papers by this authorYuhui Zang
School of Life Science and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China
Search for more papers by this authorHaiqin Lu
School of Life Science and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China
Search for more papers by this authorJie Zhu
School of Life Science and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China
Search for more papers by this authorJunchuan Qin
School of Life Science and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China
Search for more papers by this authorAbstract:
A novel recombinant dual human stem cell factor (rdhSCF) gene was constructed which consisted of a full-length hSCF cDNA plus a truncated hSCF cDNA (1–145 aa), linked by a peptide (GGGGSGGGGSGG) coding region. The rdhSCF gene was cloned into baculovirus transfer vector pAcSecG2T under the polyhedrin promoter control. Silkworm larvae infected with the recombinant virus expressed rdhSCF up to 15 800 units/mL in haemolymph. The specific activity of rdhSCF purified from the haemolymph was up to 3.0 × 106 units/mg, about 8.6 times as high as that of monomer rhSCF from Escherichia coli, and about 9.1 times as high as that of monomer rhSCF from insect cell. The binding affinity of rdhSCF to the cell surface receptor was higher than that of monomer rhSCF.
References
- 1 Williams DE, Eisenman J, Baird A, et al. Identification of a ligand for the c-kit proto-oncogene. Cell 1990; 63: 167–174.
- 2 Witte ON. Steel locus defines new multipotent growth factor. Cell 1990; 63: 5–6.
- 3 Huang EJ, Nocka KH, Beier DR, Chu T-Y, Buck J, Lahm H-W, Wellner D, Leder P, Besmer P. The hematopoietic growth factor KL is encoded by the Sl locus and is the ligand of the c-kit receptor, the product of the W locus. Cell 1990; 63: 225–233.
- 4 Zsebo KM, Williams DA, Geissier EN, et al. Stem cell factor (SCF) is encoded at the Sl locus of the mouse and is the ligand for the c-kit tyrosine kinase receptor. Cell 1990; 63: 213–224.
- 5 Hsu YR, Wu GM, Mendiaz EA, et al. The majority of stem cell factor exists as monomer under physiological conditions. J Biol Chem 1997; 272: 6406–6415.
- 6 Galli SJ, Zsebo KM, Geissler E. The kit ligand, stem cell factor. Adv Immunol 1994; 55: 1–96.
- 7 Matous JV, Langley K, Kaushansky K. Structure-function relationships of stem cell factor: an analysis based on a series of human-murine SCF chimera and the mapping of a neutralizing monoclonal antibody. Blood 1996; 88: 437–444.
- 8 Lyman D, Jacobsen W. c-kit ligand and Flt3 ligand: stem/progenitor cell factors with overlapping yet distinct activities. Blood 1998; 91: 1101–1134.
- 9 Hsu YR, Wu GM, Mendiaz EA, et al. The majority of stem cell factor exists as monomer under physiological conditions. J Biol Chem 1997; 272: 6406–6415.
- 10 Nocka KH, Levine BA, Ko JL, Burch PM, Landgraf BE, Segal R, Lobell P. Increased growth promoting but not mast cell degranulation potential of a covalent dimmer of c-kit ligand. Blood 1997; 90: 3874–3883 .
- 11 McNiece IK, Briddell RA. Stem cell factor. J Leuk Biol 1995; 58: 14–22.
- 12 Costa JJ, Demetri GD, Hayes DF, Merica EA, Menchaca DM, Galli SJ. Increased skin mast cells and urine methyl histamine in patients receiving recombinant methionyl human stem cell factor. Proc Am Assoc Cancer Res 1993; 34: 211.
- 13 Moskowitz CH, Stiff P, Gordon MS, et al. Recombinant methionyl human stem cell factor and filgrastim for peripheral blood progenitor cell mobilization and transplantation in non-Hodgkin's lymphoma patients – results of a phase I/II trial. Blood 1997; 89: 3136–3147.
- 14 Tong J, Gordon MS, Srour EF, Cooper RJ, Orazi A, McKniece I, Hoffman R. In vivo administration of recombinant methionyl human stem cell factor expands the number of human stem cells. Blood 1993; 82: 784–791.
- 15 Weaver A, Ryder D, Crowther D, Dexter TM, Testa NG. Increased numbers of long-term repopulating cells in the apheresis product of patients randomized to receive increasing doses of stem cell factor administered in combination with chemotherapy and a standard dose of granulocyte colony-stimulating factor. Blood 1999; 88: 3323–3328.
- 16 Jinag X, Gurel O, et al. Structure of the active core of human stem cell factor and analysis of binding to its receptor kit. EMBO 2000; 19: 3192–3203.
- 17 Zhang Z, Zhang R, Joachimiak A, Schlessinger J, Kong X-P. Crystal structure of human stem cell factor: implication for stem cell factor receptor dimerization and activation. PANS 2000; 97: 7732–7737.
- 18 Felgner PL, Gadek TR, Holm M, Roman R, Chan HW, Wenz M, Northrop JP, Ringold GM, Danielsen M. Lipofectin: a highly efficient, liquid-mediated DNA-transfection procedure. Proc Natl Acad Sci USA 1987; 84: 7413–7417.
- 19 Summers MD, Smith GE. A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures. Texas, USA Texas Agriculture Experiment Station, 1987, Bulletin No. 1555.
- 20 Han JH, Yan XH, Zhu J, Zhi XY, Zang YH, Shen BF, Qin JC. Expression of a novel recombinant dual human stem cell factor in insect cells. Protein Expression and Purification 2003; 31: 311–317.
- 21 Turner AM, Bennett LG, Lin NL, et al. Identification and characterization of a soluble c-kit receptor produced by human hematopoietic cell lines. Blood 1995; 85: 2052.
- 22 Broudy VC, Kovach NL, Bennett LG, Lin N, Jacobsen FW, Kidd PG. Human umbilical vein endothelial cells display high affinity c-kit receptors and produce a soluble form of the c-kit receptor. Blood 1994; 83: 2145–2152.
- 23 Munson PJ, Rodbard D. A versatile computerized approach for characterization of ligand-binding system. Anal Biochem 1980; 107: 220–239.
- 24 Peng R, Yu Z, Zang Y, Qin J. High level expression of a mutant (K151E, R154G) of single chain urokinase-type plasminogen activator in silkworm. Biotechnol Lett 1999; 21: 979–985.
- 25 Martin FH, Suggs SV, Langley KE, et al. Primary structure and functional expression of rat and human stem cell factor DNAs. Cell 1990; 63: 203–211.
- 26 Langley KE, Wypych J, Mendiaz EA, et al. Purification and characterization of soluble forms of human and rat stem cell factor recombinantly expressed by Escherichia coli and by Chinese hamster ovary cells. Arch Biochem Biophys 1992; 295: 21.
- 27 Broudy VC, Smith FO, Lin N, Zsebo KM, Egrie J, Bernstein ID. Blasts from patients with acute myelogenous leukemia express functional receptors for stem cell factor. Blood 1992; 80: 60–67.
- 28 Turner AM, Zsebo KM, Martin F, Jacobsen FW, Bennett LG, Broudy VC. Nonhematopoietic tumor cell lines express stem cell factor and display c-kit receptors. Blood 1992; 80: 374–381.
- 29 Lemmon MA, Pinchasi D, Zhou M, Lax I, Schlessinger J. Kit receptor dimerization is driven by bivalent binding of stem cell factor. J Biol Chem 1997; 272: 6311–6317.
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