Characterization of monomeric and homodimeric forms of osteoclastogenesis inhibitory factor.
Osteoclastogenesis inhibitory factor (OCIF) is present naturally as two molecular forms, a monomer and a homodimer. The two forms of recombinant human OCIF (rOCIF) produced by Chinese hamster ovary (CHO) cells were purified to homogeneity. Determination of the C-terminal amino-acid sequences of the two forms of rOCIF revealed that the monomeric rOCIF lacked several amino acids including Cys379, which is involved in the intermolecular disulfide bond, in its C-terminal region. The two forms of rOCIF were indistinguishable in stability, sialic acid content, and specific activity in inhibition of osteoclastogenesis. In contrast, the homodimeric rOCIF was stronger in heparin-binding ability than the monomeric rOCIF. The homodimeric rOCIF was significantly shorter in initial half-life and smaller in AUC value in rats than the monomeric rOCIF, but exerted more potent biological activity in reducing the calcium concentration in serum of rats than did the monomeric rOCIF.
Tomoyasu
A
A
Research Institute of Life Science, Snow Brand Milk Products Co., Ltd., Tochigi, Japan.
Goto
M
M
Fujise
N
N
Mochizuki
S
S
Yasuda
H
H
Morinaga
T
T
Tsuda
E
E
Higashio
K
K
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002118:Calcium / Q000097:blood; D006224:Cricetinae; D019281:Dimerization; D004220:Disulfides / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry* / Q000493:pharmacokinetics; D006493:Heparin / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid / Q000032:analysis; D053244:Osteoprotegerin; D011485:Protein Binding / Q000502:physiology; D018160:Receptors, Cytoplasmic and Nuclear; D018124:Receptors, Tumor Necrosis Factor; D011994:Recombinant Proteins / Q000737:chemistry; D017421:Sequence Analysis
D004220:Disulfides; D006023:Glycoproteins; D053244:Osteoprotegerin; D018160:Receptors, Cytoplasmic and Nuclear; D018124:Receptors, Tumor Necrosis Factor; D011994:Recombinant Proteins; C506030:TNFRSF11B protein, human; D006493:Heparin; D019158:N-Acetylneuraminic Acid; D002118:Calcium