Human macrophage colony stimulating factor (HM-CSF) expressed in baculovirus infected insect cells is biologically active in its monomeric form

Biochem Mol Biol Int. 1995 Apr;35(4):773-9.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Base Sequence
  • Binding, Competitive
  • Cells, Cultured
  • DNA, Complementary / genetics
  • Genetic Vectors
  • Humans
  • Insecta
  • Macrophages / metabolism
  • Mice
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Occlusion Body Matrix Proteins
  • Point Mutation
  • Promoter Regions, Genetic / genetics
  • Receptor, Macrophage Colony-Stimulating Factor / biosynthesis
  • Receptor, Macrophage Colony-Stimulating Factor / genetics*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / pharmacology*
  • Viral Proteins / genetics
  • Viral Structural Proteins

Substances

  • DNA, Complementary
  • Occlusion Body Matrix Proteins
  • Recombinant Proteins
  • Viral Proteins
  • Viral Structural Proteins
  • polyhedrin protein, Nucleopolyhedrovirus
  • Receptor, Macrophage Colony-Stimulating Factor