Purification and subunit structure of hepatocyte growth factor from rat platelets.
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 C4 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).
Nakamura
T
T
Institute for Enzyme Research, University of Tokushima, Japan.
Nawa
K
K
Ichihara
A
A
Kaise
N
N
Nishino
T
T
D000818:Animals; D001792:Blood Platelets / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D015850:Interleukin-6; D046911:Macromolecular Substances; D008970:Molecular Weight; D011506:Proteins / Q000302:isolation & purification*; D051381:Rats
D015850:Interleukin-6; D046911:Macromolecular Substances; D011506:Proteins