Purification and subunit structure of hepatocyte growth factor from rat platelets

FEBS Letters

Nishino, Tomoyoshi¹  Kaise, Nobuko¹  Nawa, Katsuhiko²  Ichihara, Akira²  Nakamura, Toshikazu² 

[1] Research Laboratories, Otsuka Pharmaceutical Co. Ltd, Tokushima 771-01, Japan;Institute for Enzyme Research, University of Tokushima, Tokushima 770, Japan

关键词: Hepatocyte growth factor;    Primary cultured hepatocyte;    Platelet;    Heterodimer;   

DOI  :  10.1016/0014-5793(87)80475-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 beat- 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).

【 授权许可】

Unknown