Structural characterization of natural and recombinant human granulocyte colony-stimulating factors.
Granulocyte colony-stimulating factor (G-CSF) is a glycoprotein which stimulates predominantly neutrophilic granulocyte colony formation in mammals. Natural human G-CSF (hG-CSF) and recombinant hG-CSF produced in Chinese hamster ovary (CHO) cells transfected with the cDNA clone for hG-CSF have been purified to apparent homogeneity for structural and biological comparison. The amino acid sequence of recombinant hG-CSF, composed of 174 amino acid residues, was identical with that of natural hG-CSF and also with the sequence predicted from the cDNA. Both forms of hG-CSF have a free Cys-17 and two intramolecular disulfide linkages, between Cys-36 and Cys-42, and between Cys-64 and Cys-74. The O-glycosylation occurred at Thr-133 in both hG-CSFs. Similar CD spectra were obtained for both hG-CSFs. Additionally, both forms showed almost the same biological activities determined by in vitro colony-forming assay and in vivo assay. It is thus concluded that the recombinant hG-CSF is indistinguishable from its natural counterpart and that the former is valuable for more detailed characterization and clinical use.
Fuji-Gotemba Research Laboratories, Chugai Pharmaceutical Co., Ltd., Shizuoka.
N
N
Kubota
T
T
Orita
K
K
Hattori
M
M
Oh-eda
N
N
Ochi
T
T
Yamazaki
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002478:Cells, Cultured; D002942:Circular Dichroism; D003114:Colony-Forming Units Assay; D003115:Colony-Stimulating Factors / Q000032:analysis* / Q000494:pharmacology; D006224:Cricetinae; D003412:Cricetulus; D004220:Disulfides / Q000032:analysis; D016179:Granulocyte Colony-Stimulating Factor; D006801:Humans; D006868:Hydrolysis; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000032:analysis / Q000494:pharmacology; D012697:Serine Endopeptidases
D000596:Amino Acids; D003115:Colony-Stimulating Factors; D004220:Disulfides; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase