Purification and characterization of the biologically active human truncated macrophage colony-stimulating factor expressed in Saccharomyces cerevisiae

Biol Chem Hoppe Seyler. 1993 Sep;374(9):903-8. doi: 10.1515/bchm3.1993.374.7-12.903.

Abstract

A human truncated macrophage colony-stimulating factor (M-CSF) cDNA encoding amino acid residues from 3 to 149 of the native M-CSF was isolated by using the polymerase chain reaction. When introduced into Saccharomyces cerevisiae by a general secretion vector pVT 102u/alpha, it directs the expression of the biologically active dimeric form of M-CSF. Through the 3 stages of purification, i.e. concentration by DEAE-cellulose column chromatography, hydrophobic chromatography on phenyl-sepharose and Mono Q fast protein liquid chromatography, the recombinant truncated M-CSF was purified as to exhibit a specific activity of 1.02 x 10(7) units/mg of protein. SDS-PAGE of this purified truncated M-CSF showed that its apparent molecular mass is 22 kDa under reducing conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Chromatography, Liquid
  • DNA, Complementary / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Macrophage Colony-Stimulating Factor / biosynthesis
  • Macrophage Colony-Stimulating Factor / chemistry
  • Macrophage Colony-Stimulating Factor / genetics
  • Macrophage Colony-Stimulating Factor / isolation & purification*
  • Molecular Sequence Data
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Sequence Analysis, DNA

Substances

  • DNA, Complementary
  • Recombinant Proteins
  • Macrophage Colony-Stimulating Factor