Purification and subunit structure of hepatocyte growth factor from rat platelets

Abstract

A hepatocyte growth factor (HGF) that stimulates DNA synthesis of adult rat hepatocytes in primary culture was purified as a homogeneous material from platelets of 1000 rats by a four-step procedure: stimulation of its release from platelets by thrombin, cation-exchanger fast protein liquid chromatography (FPLC) on a Mono S column, heparin-Sepharose CL-6B chromatography, and reverse-phase HPLC on a C 4 column. The purified HGF stimulated DNA synthesis of adult rat hepatocytes in primary culture at 1 ng/ml and was maximally effective at 5 ng/ml, being about twice as potent as EGF at this concentration. HGF did not stimulate DNA synthesis of Swiss 3T3 cells. It was found to be a heat-and acid-labile protein that was inactivated by reduction with dithiothreitol. The purified HGF had a molecular mass of 82 kDa, as estimated by SDS-PAGE, and was found to be a heterodimer which dissociated into a large subunit of 69 kDa and a small one of 34 kDa by SDS-PAGE under reducing conditions. These biological and chemical properties showed that HGF was not identical with any known growth factors, including platelet-derived growth factor (PDGF).

Concomitant expression of hepatocyte growth factor (HGF), HGF activator and c‐met genes in human glioma cells in vitro

“…HGF is a multifunctional, heparin-binding glycoprotein initially identified as a potent mitogen for hepatocytes [1,2]. The c-met proto-oncogene product, a membrane-bound tyrosine kinase, was demonstrated to represent the high affinity HGF receptor suggesting the roles of HGF in tumor progression [6].…”

“…These processes are often mediated by polypeptide growth factors acting in either a paracrine or autocrine fashion. Hepatocyte growth factor (HGF) is a pleiotropic factor initially identified as a polypeptide growth factor for hepatocytes [1,2], and is indistinguishable from scatter factor (SF), a motility factor [3][4][5][6]. HGF functions as a mitogen for a variety of cell types, as a morphogen and as a motogen for some epithelial cells [6].…”

Hepatocyte growth factor/scatter factor (HGF/SF) affects proliferation and migration of myeloid leukemic cells

“…[1][2][3][4][5] HGF/SF is produced by a variety of mesenchymal cells as a single chain pro-HGF/SF, which is enzymatically, cleaved into biologically active HGF/SF consisting of an ␣-and a ␤-chain. [6][7][8][9][10] The receptor for HGF/SF is encoded by the MET proto-oncogene, c-MET, which is a cell surface tyrosine kinase receptor also consisting of an extracellular ␣-and a transmembrane ␤-chain. 11 The ␤-chain contains the tyrosine kinase domain, as well as sites for tyrosine autophosphorylation.…”

Phorbol ester‐induced secretion of human hepatocyte growth factor by human skin fibroblasts and its inhibition by dexamethasone

“…which stimulates DNA synthesis on adult rat hepatocytes in primary culture, was purified from the plasma of patients with fulminant hepatic failure [I], rat platetcs [2], rabbit serum [3], normal human plasma [3], and conditioned human embryonic lung fibroblast medium [4], HGF is a heterodimcr which consists of a heavy chain of about 60,000 Da, and a light chain of about 35,000 Da. linked together, probably by a single disulfide bond [ I,-41.…”