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Two histidine residues are essential for ribonuclease T/sub 1/ activity as is the case for ribonuclease

Journal Article · · Biochemistry; (United States)
OSTI ID:5381682
Ribonuclease T/sub 1/ (RNase T/sub 1/, EC 3.1.27.3) is a guanosine-specific ribonuclease that cleaves the 3',5'-phosphodiester linkage of single-stranded RNA. It is assumed that the reaction is generated by concerted acid-base catalysis between residues Glu-58 and His-92 or His-40. From the results of chemical modification and NMR studies, it appeared that the residue Glu-58 was indispensable for nucleolytic activity. However, the authors have recently demonstrated that Glu-58 is an important but not an essential residue for catalytic activity, using the methods of genetic engineering to change Glu-58 to Gln-58 etc. In the present paper, the authors report that mutants of RNase T/sub 1/ with residue Ala-40 or Ala-92 have almost no activity, while mutants that contain Ala-58 retain considerable activity. These results show that the two histidine residues, His-40 and His-92, but not Glu-58, are indispensable for the catalytic activity of the enzyme. They propose a revised reaction mechanism in which two histidine residues play a major role, as they do in the case of RNase A.
Research Organization:
Osaka Univ. (Japan)
OSTI ID:
5381682
Journal Information:
Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 26:26; ISSN BICHA
Country of Publication:
United States
Language:
English