Purification and characterization of hepatocyte growth factor (HGF)-converting enzyme: activation of pro-HGF.
Hepatocyte growth factor (HGF) is a heterodimer protein, derived from an inactive single-chain precursor (pro-HGF) by the proteolytic cleavage at Arg-Val site. Using a fluorogenic substrate Ac-Lys-Thr-Lys-Gln-Leu-Arg-MCA corresponding to the sequence around the cleavage site of pro-HGF, HGF-converting enzyme was purified from fetal bovine serum. The enzyme is a heterodimer molecule with an apparent molecular weight of about 90-kDa and is composed of a 65-kDa heavy-chain and a 32-kDa light-chain. The enzyme belongs to the serine-protease family and has an optimal pH around 8. The enzyme preferentially cleaved MCA-substrates containing the processing site of pro-HGF. The purified enzyme converted pro-HGF to a two-chain mature form of HGF. The enzyme-treated pro-HGF had mitogenic activity on primary cultured hepatocytes. Thus, the enzyme is likely to be involved in pro-HGF activation in vivo. The enzyme activity in rat serum was 9-fold higher than that in the plasma. This, together with the heterodimeric structure of the enzyme, suggests that the HGF-converting enzyme is activated by another protease in response to a trigger such as blood coagulation or tissue injury.
Mizuno
K
K
Department of Biology, Faculty of Science, Kyushu University, Fukuoka, Japan.
Tanoue
Y
Y
Okano
I
I
Harano
T
T
Takada
K
K
Nakamura
T
T
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002417:Cattle; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006224:Cricetinae; D004247:DNA / Q000096:biosynthesis; D017228:Hepatocyte Growth Factor / Q000096:biosynthesis* / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000096:biosynthesis / Q000302:isolation & purification* / Q000378:metabolism*; D013379:Substrate Specificity; D014162:Transfection
D009842:Oligopeptides; D011498:Protein Precursors; D011994:Recombinant Proteins; C086013:pro-hepatocyte growth factor; D017228:Hepatocyte Growth Factor; D004247:DNA; D012697:Serine Endopeptidases; C086012:hepatocyte growth factor-converting enzyme