v-Ha-Ras insertion/deletion mutants with reduced protease-inhibitory activity have no transforming activity.
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. Ki 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.
Sawada
T
T
Division of Biochemistry, Chiba Cancer Center Research Institute, Japan.
Sakiyama
S
S
Hiwasa
T
T
D016475:3T3 Cells; D000818:Animals; D002401:Cathepsin B / Q000037:antagonists & inhibitors; D056668:Cathepsin L; D002403:Cathepsins / Q000037:antagonists & inhibitors; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D003546:Cysteine Endopeptidases; D010450:Endopeptidases; D004926:Escherichia coli / Q000235:genetics*; D017353:Gene Deletion; D011905:Genes, ras / Q000235:genetics*; D051379:Mice; D009154:Mutation; D010206:Papain / Q000037:antagonists & inhibitors; D011480:Protease Inhibitors / Q000494:pharmacology*; D013329:Structure-Activity Relationship
D011480:Protease Inhibitors; D002403:Cathepsins; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; D056668:Cathepsin L; C534278:Ctsl protein, mouse; D010206:Papain