Influence of gelatin complexation on cell proliferation activity and proteolytic resistance of basic fibroblast growth factor.
The objective of this study is to investigate the influence of gelatin complexation on the biological activity of basic fibroblast growth factor (bFGF) and its resistance to trypsin digestion. When bFGF was mixed at 37 degrees C with acidic gelatin with an isoelectric point (IEP) of 5.0, the activity to promote in vitro proliferation of BHK cells became lower compared with that of free bFGF, in contrast to mixing with the basic gelatin with an IEP of 9.0. A maximum reduction in the bFGF activity was observed for the bFGF-gelatin complex prepared at a mixing molar ratio of 1/1. The bFGF activity of cell proliferation reduced at the initial period after mixing with the acidic gelatin at 37 degrees C, followed by no substantial change. Complexation with the acidic gelatin at 4 degrees C had no influence on the bFGF activity, irrespective of the bFGF/gelatin ratio and complexation time. The biological activity of bFGF was reduced by the trypsin treatment, but the reduced extent was suppressed through gelatin complexation at 37 degrees C. In an electrophoresis study, the protective effect of gelatin complexation on the trypsin digestion was also confirmed in terms of the molecular weight loss. It is possible that the complexing gelatin covers bFGF molecules, resulting in suppression of their interaction with the cell surface receptor as well as protection from their enzymatic attack.
Tabata
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Institute for Frontier Medical Sciences, Kyoto University, Japan. yasuhiko@frontier.kyoto-u.ac.jp
Ishii
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Muniruzzaman
M
M
Hirano
Y
Y
Ikada
Y
Y
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry / Q000378:metabolism*; D005780:Gelatin / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007526:Isoelectric Point; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D013696:Temperature; D013997:Time Factors
D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; D005780:Gelatin