v‐Ha‐Ras insertion/deletion mutants with reduced protease‐inhibitory activity have no transforming activity

FEBS Letters

Sakiyama, Shigeru¹  Sawada, Toshie¹  Hiwasa, Takaki¹ 

[1] Division of Biochemistry, Chiba Cancer Center Research Institute, 666-2 Nitona-cho, Chuo-ku, Chiba 260, Japan

关键词: v-Ha-ras;    Oncogene product;    Protease inhibitor;    Cathepsin;    GTP binding;    DTT;    dithiothreitol;    E. coli;    Escherichia coli;    GAP;    GTPase activating protein;    MES;    2-(N-morpholino)ethanesulfonic acid;    Ras;    ras oncogene product;    Z-Arg-Arg-MCA;    7-(benzyloxycarbonyl-arginyl-arginyl-amino)-4-methylcoumarin;    Z-Phe-Arg-MCA;    7-(benzyloxycarbonyl-phenylalanyl-arginyl-amino)-4-methylcoumarin;   

DOI  :  10.1016/0014-5793(93)80532-Y

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have purified 26 insertion/deletion mutants of v-Ha-ras oncogene products produced by Escherichia coli and investigated their protease-inhibitory activity toward papain and cathepsins B and L. K _i values for papain were relatively similar among the mutants, however, those for cathepsins B and L varied up to 10-fold. Among them, four mutants, 1–48 LIR 54–189, 1–110 LIS 112–189, 1–130 PDQ 146–189 and 1–155 LIR 166–189, showed significant reduction in the inhibitory activity toward cathepsin L and these four mutants have lost transforming activity toward NIH3T3 mouse fibroblasts. However, some other mutants also showed no transforming activity in spite of possession of the potent protease-inhibitory activity, suggesting that the protease-inhibitory activity of Ras might be necessary but not sufficient for its biological activity.

【 授权许可】

Unknown