title: Characterization of recombinant-derived granulocyte-colony stimulating factor (G-CSF).
abstract: Human granulocyte colony-stimulating factor (G-CSF), and a mutant having a Ser for Cys substitution at residue 18 were produced in Escherichia coli strain W3110. About 60 mg of pure protein was obtained from 50 g of wet cells with a recovery of about 20%. The proteins were characterized physically and chemically, including determination of disulphide bonds, which were found to exist between residues 37-43 and 65-75. Cys-18 is not involved in disulphide bond formation and was substituted by Ser with no effects on gross protein conformation or biological activity. Both the wild-type and the mutant recombinant-derived proteins, although not glycosylated, possess colony-stimulating activities. In a bioassay using the murine myelomonocytic leukaemic cell line WEH1 3B D+, activities were obtained which were similar to those of natural G-CSF and of a glycosylated recombinant-derived human G-CSF produced in monkey cells.
authors: Biogen S.A., Geneva, Switzerland.
authors: P
authors: P
authors: Wingfield
authors: R
authors: R
authors: Benedict
authors: G
authors: G
authors: Turcatti
authors: B
authors: B
authors: Allet
authors: J J
authors: JJ
authors: Mermod
authors: J
authors: J
authors: DeLamarter
authors: M G
authors: MG
authors: Simona
authors: K
authors: K
authors: Rose
mesh_terms: D000595:Amino Acid Sequence; D003115:Colony-Stimulating Factors / Q000096:biosynthesis; D004274:DNA, Recombinant / Q000378:metabolism; D004220:Disulfides / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D016179:Granulocyte Colony-Stimulating Factor; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000096:biosynthesis
chemical_list: D003115:Colony-Stimulating Factors; D004274:DNA, Recombinant; D004220:Disulfides; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor