[Relationship between biological activity and heparin-affinity sites of recombinant human basic fibroblast growth factor CS23 mutein].
A recombinant human basic fibroblast growth factor CS23 mutein (rhbFGF-CS23) obtained from Escherichia coli cells has proliferation-stimulating activity for a fetal bovine heart endothelial cell line, ATCC CRL 1395 (biological activity), and strong affinity for heparin (heparin-affinity) similarly to the natural human basic fibroblast growth factor. Plural species having different kinds of heparin-affinity were formed by acetylation of rhbFGF-CS23 with acetic anhydride. To clarify the relationship between the sites with heparin-affinity and the biologically active sites, we have investigated the acetylation sites by peptide mapping and the biological activity of the acetylated species. Consequently, the sites with heparin-affinity in rhbFGF-CS23 are found to be Lys26, Lys119, Lys125, Lys129, and Lys135 in the primary structure, and these sites with heparin-affinity except Lys26 are also clarified to be very important to retain the biological activity of the factor.
Hiroshima
T
T
Drug Analysis & Pharmacokinetics Research Laboratories, Pharmaceutical Development Division, Takeda Chemical Industries, Ltd., Osaka, Japan.
Higashi
A
A
Kusumoto
S
S
Nishi
K
K
Nishijima
K
K
D000107:Acetylation; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D005346:Fibroblast Growth Factors / Q000378:metabolism / Q000502:physiology; D006493:Heparin / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011994:Recombinant Proteins / Q000378:metabolism
D011994:Recombinant Proteins; C065198:protein CS23; D005346:Fibroblast Growth Factors; D006493:Heparin