Activation of hepatocyte growth factor by proteolytic conversion of a single chain form to a heterodimer.
Hepatocyte growth factor (HGF) is a heterodimeric protein consisting of a heavy chain and a light chain held by a disulfide bond. These chains are produced by endoproteolytic processing from a single chain precursor. In this study, we examined whether the processing is a prerequisite for the mitogenic activity of HGF on hepatocytes in primary culture. Single chain HGF was proteolytically converted to the heterodimeric form during incubation with hepatocytes and was as mitogenic as the heterodimeric form. When the conversion was inhibited by serine-protease inhibitors, the mitogenic activity of single chain HGF was markedly reduced. Furthermore, a mutant resistant to the proteolytic processing, which was prepared by in vitro mutagenesis, completely lost the mitogenic activity. From these results, we concluded that the single chain form of HGF is endoproteolytically processed by a serine-protease and that this processing is a prerequisite for the mitogenic activity of HGF.
Naka
D
D
Biosciences Laboratory, Mitsubishi Kasei Corporation, Yokohama, Japan.
Ishii
T
T
Yoshiyama
Y
Y
Miyazawa
K
K
Hara
H
H
Hishida
T
T
Kidamura
N
N
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D004247:DNA / Q000096:biosynthesis; D004591:Electrophoresis, Polyacrylamide Gel; D017228:Hepatocyte Growth Factor / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D008099:Liver / Q000166:cytology / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D011499:Protein Processing, Post-Translational; D019859:Proto-Oncogene Proteins c-met; D051381:Rats; D017208:Rats, Wistar; D011956:Receptors, Cell Surface / Q000378:metabolism; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology
D011956:Receptors, Cell Surface; D015842:Serine Proteinase Inhibitors; D017228:Hepatocyte Growth Factor; D004247:DNA; D019859:Proto-Oncogene Proteins c-met