期刊
FEBS LETTERS
卷 466, 期 1, 页码 87-90出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/S0014-5793(99)01753-6
关键词
mitogillin; restrictocin; ribotoxin; specificity; targeted toxin; alpha-sarcin/ricin loop
Mitogillin and related fungal ribotoxins are small basic ribonucleolytic proteins that inhibit protein synthesis by specifically hydrolyzing a single phosphodiester bond in the universally conserved alpha-sarcin/ricin loop (SRL) of large subunit ribosomal RNAs. It was previously shown that mitogillin is a natural derivative of a T1/U2-like ribonuclease with inserted domains that are involved in target selection and specificity. Site-directed mutagenesis was used to substitute single amino acids in the previously identified functional domains Alal-Tyr24 (B1-L1-B2 domain) and Lys106-Lys113 (L4 region). Examination of the activities of the mutants in the digestion of polyinosinic acid (a ribonuclease substrate) and specific cleavage of the SRL shows that Asn7Ala and Lys111Gln substitutions lead to altered ribonuclease activity and diminished substrate specificity consistent with the proposed functions of these domains. (C) 2000 Federation of European Biochemical Societies.
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