期刊
PROTEIN SCIENCE
卷 11, 期 10, 页码 2522-2525出版社
COLD SPRING HARBOR LAB PRESS
DOI: 10.1110/ps.0216702
关键词
ribonuclease Sa; net charge; charge reversal mutants; cytotoxicity; v-ras-transformed fibroblasts
资金
- FIC NIH HHS [TW01058] Funding Source: Medline
- NIGMS NIH HHS [R01 GM037039] Funding Source: Medline
Ribonuclease Sa (pI = 3.5) from Streptomyces aureofaciens and its 3K (D1K, D17K, E41K) (pI = 6.4) and 5K (3K + D25K, E74K) (pI = 10.2) mutants were tested for cytotoxicity. The 5K mutant was cytotoxic to normal and v-ras-transformed NIH3T3 mouse fibroblasts, but RNase Sa and 3K were not. The structure. stability, and activity of the three proteins are comparable, but the net charge at pH 7 increases from -7 for RNase Sa to -1 for 3K and to +3 for 5K. These results suggest that a net positive charge is a key determinant of ribonuclease cytotoxicity. The cytotoxic 5K mutant preferentially attacks v-ras-NIH3T3 fibroblasts, suggesting that mammalian cells expressing the ras-oncogene are potential targets for ribonuclease-based drugs.
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