Analysis of deleted variant of hepatocyte growth factor by alanine scanning mutagenesis: identification of residues essential for its biological function and generation of mutants with enhanced mitogenic activity on rat hepatocytes

FEBS Letters

Kinosaki, Masahiko¹  Yamaguchi, Kyoji¹  Murakami, Akihiko¹  Ueda, Masatsugu¹  Higashio, Kanji¹  Morinaga, Tomonori¹ 

[1] Research Institute of Life Science, Snow Brand Milk Products Co. Ltd., 519 Ishibashi-machi, Shimotsuga-gun, Tochigi 329-0512, Japan

关键词: Hepatocyte growth factor;    Deleted variant of hepatocyte growth factor;    Alanine scanning mutagenesis;    dHGF;    deleted variant of hepatocyte growth factor;    HGF;    hepatocyte growth factor;    ELISA;    enzyme-linked immunosorbent assay;    IMDM;    Iscove's modified Dulbecco's medium;    FPLC;    fast protein liquid chromatography;    FBS;    fetal bovine serum;    DMEM;    Dulbecco's modified Eagle medium;    PBS;    phosphate-buffered saline;    EC50;    dose required for 50% maximal cell proliferative response;   

DOI  :  10.1016/S0014-5793(98)00971-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

To understand the structure-function relationship of hepatocyte growth factor (HGF) in more detail, we analyzed one of the other forms of HGF, deleted variant of HGF (dHGF), by alanine scanning mutagenesis. We show here that there are at least four sites important for dHGF to stimulate DNA synthesis in cultured adult rat hepatocytes, and that the residues of HGF essential for exerting its biological activity are not identical to those of dHGF. In addition, two mutants showed a decrease (approximately three-fold) in EC₅₀ compared with wild-type dHGF in an assay of mitogenic activity on rat hepatocytes.

【 授权许可】

Unknown