Identification of binding domains for basic fibroblast growth factor in proteoglycan macrophage colony-stimulating factor.
We recently demonstrated that proteoglycan macrophage colony-stimulating factor (PG-M-CSF) binds basic fibroblast growth factor (bFGF) and neutralizes the biological activity of bFGF. In this study, we identified the binding sites of PG-M-CSF for bFGF. We examined the binding of bFGF to overlapping 12-mer peptides with the sequence of the putative binding region. High affinity binding was detected at two peaks; one consisted of the three adjacent peptides, 212-223, 213-224 and 214-225 and the other, of the three adjacent peptides, 246-257, 247-258 and 248-259. The synthetic peptide (212VDPGSAKQRPPRST225) did not inhibit bFGF binding to another peptide (246PQPRPSVGAFNPGM259), and vice versa. However, both peptides inhibited the bFGF-induced but not platelet-derived growth factor-induced stimulation of DNA synthesis in murine Balb/c 3T3 cells.
Suzu
S
S
Biochemical Research Laboratory, Morinaga Milk Industry Co., Ltd., Kanagawa, Japan.
Kimura
F
F
Matsumoto
H
H
Yamada
M
M
Hashimoto
K
K
Shimamura
S
S
Motoyoshi
K
K
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D002455:Cell Division / Q000187:drug effects; D006224:Cricetinae; D004247:DNA / Q000096:biosynthesis; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D016173:Macrophage Colony-Stimulating Factor / Q000737:chemistry* / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D011509:Proteoglycans / Q000737:chemistry* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014162:Transfection
D010446:Peptide Fragments; D011509:Proteoglycans; D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; D016173:Macrophage Colony-Stimulating Factor; D004247:DNA