Human macrophage colony stimulating factor (HM-CSF) expressed in baculovirus infected insect cells is biologically active in its monomeric form.
hM-CSF was reported to have biological activity only in a dimeric form. Using oligonucleotide-directed mutagenesis of hM-CSF (1-149aa) cDNA, we have substituted Ser31 for Cys31 which forms intermolecular disulfide bond in native hM-CSF. The mutant hM-CSF cDNA was expressed in insect BmN cells using baculovirus as a vector under the control of polyhedrin promoter. Biological activity analysis and radioligand receptor assay both showed that there was little difference between the mutant hM-CSF and the native dimeric hM-CSF. These results strongly support that the biologically active human M-CSF in its monomeric form can be expressed in recombinant baculovirus infected insect cells.
Qiu
P
P
Department of Biochemistry, Nanjing University, P.R. China.
Li
J
J
Qin
J
J
Xi
T
T
Zhu
D
D
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D001667:Binding, Competitive; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D007313:Insecta; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D000077183:Occlusion Body Matrix Proteins; D017354:Point Mutation; D011401:Promoter Regions, Genetic / Q000235:genetics; D016186:Receptor, Macrophage Colony-Stimulating Factor / Q000096:biosynthesis / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000494:pharmacology*; D014764:Viral Proteins / Q000235:genetics; D015678:Viral Structural Proteins
D018076:DNA, Complementary; D000077183:Occlusion Body Matrix Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus; D016186:Receptor, Macrophage Colony-Stimulating Factor