Interaction of hepatocyte growth factor with gelatin as the carrier material.
The objective of this study was to physicochemically investigate the interaction between hepatocyte growth factor (HGF) and acidic gelatin compared with that between HGF and basic gelatin or heparin. Gelatin- or heparin-immobilized agarose beads were prepared and HGF interaction with them was evaluated by Scatchard binding assay. The dissociation constant of HGF with the acidic gelatin was about 2-3 orders of magnitude higher than that of heparin. The cell proliferation assay revealed that the proliferation promotion activity of HGF complexed with the acidic gelatin was detected, although it was lower than that of original HGF. The ability of HGF to enhance the cell proliferation was reduced by the trypsin treatment, although the extent of the reduction was significantly suppressed by HGF complexation with acidic gelatin. Electrophoresis experimentally confirmed enhanced resistance to the molecular mass loss of HGF by gelatin complexation. Moreover, the recognized level of an antibody to HGF was reduced by the complexation with the acidic gelatin, indicating that the acidic gelatin is present around HGF molecules. It is possible that the HGF molecule is covered with the acidic gelatin, resulting in protection from enzymatic digestion.
Ozeki
Makoto
M
Institute of Frontier Medical Sciences, Kyoto University, Shogoin, Sakyo-ku, Japan.
Tabata
Yasuhiko
Y
D000906:Antibodies / Q000276:immunology; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D016503:Drug Delivery Systems / Q000379:methods*; D004586:Electrophoresis; D004797:Enzyme-Linked Immunosorbent Assay; D005780:Gelatin / Q000737:chemistry* / Q000378:metabolism; D006493:Heparin / Q000737:chemistry; D017228:Hepatocyte Growth Factor / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015394:Molecular Structure; D009994:Osmolar Concentration; D012996:Solutions; D014357:Trypsin / Q000378:metabolism
D000906:Antibodies; D012996:Solutions; D017228:Hepatocyte Growth Factor; D005780:Gelatin; D006493:Heparin; D014357:Trypsin