Purification and characterization of hepatocyte growth factor (HGF)-converting enzyme: activation of pro-HGF

Abstract

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.