Effects of herbimycin A and various SH-reagents on p60v-src kinase activity in vitro.
Herbimycin A is an antiobiotic which reverses transformation caused by src family oncogenes. It inactivates p60v-src in vitro, possibly by binding to reactive SH-group(s) of the kinase. We examined effects of various SH-reagents on p60v-src and observed that N-[p-(2-benzimidazolyl)phenyl]maleimide (BIPM) or N-(9-acridinyl)maleimide (NAM) were potent inactivators of the kinase, whereas N-ethylmaleimide (NEM) required high concentrations, and iodoacetamide was totally ineffective in reducing the kinase activity. Pretreatment of p60v-src immune-complex with NEM and iodoacetamide, however, protected the kinase from inactivation by herbimycin A, BIPM, and NAM. The results suggest that SH-group(s) to which herbimycin A binds is not essential for the kinase activity, but is positioned in the vicinity of the active center.
Fukazawa
H
H
Department of Antibiotics, National Institute of Health, Tokyo, Japan.
Mizuno
S
S
Uehara
Y
Y
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D001358:Avian Sarcoma Viruses / Q000235:genetics; D016227:Benzoquinones; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D005033:Ethylmaleimide / Q000494:pharmacology; D007460:Iodoacetamide / Q000494:pharmacology; D007700:Kinetics; D047029:Lactams, Macrocyclic; D051379:Mice; D015688:Oncogene Protein pp60(v-src) / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011809:Quinones / Q000494:pharmacology*; D017828:Rifabutin / Q000031:analogs & derivatives; D013439:Sulfhydryl Reagents / Q000494:pharmacology*
D000903:Antibiotics, Antineoplastic; D016227:Benzoquinones; D047029:Lactams, Macrocyclic; D011809:Quinones; D013439:Sulfhydryl Reagents; D017828:Rifabutin; C020754:herbimycin; D011505:Protein-Tyrosine Kinases; D015688:Oncogene Protein pp60(v-src); D005033:Ethylmaleimide; D007460:Iodoacetamide