Identification by site-directed mutagenesis of amino acids in the subsite of bovine pancreatic ribonuclease A
| Autor: | Henny J. Eggelte, Stuart Harbron, Brian R. Rabin, Christopher John Taylorson, Antonio Tarragona-Fiol, John M. Ward, E. Sanchez |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
chemistry.chemical_classification
biology Guanosine Bioengineering Protein engineering Bovine pancreatic ribonuclease Biochemistry Amino acid chemistry.chemical_compound chemistry biology.protein Pancreatic ribonuclease Ribonuclease Site-directed mutagenesis Molecular Biology Nucleoside Biotechnology |
| Zdroj: | "Protein Engineering, Design and Selection". 6:901-906 |
| ISSN: | 1741-0134 1741-0126 |
| DOI: | 10.1093/protein/6.8.901 |
| Popis: | In addition to hydrolysing RNA, bovine pancreatic ribonuclease splits esters of pyrimidine nucleoside 3'-phosphates, including dinucleotides. For a series of 3':5'-linked dinucleotides of general structure CpN, where N is a 5' linked nucleoside, kcat for the release of N varies enormously with the precise structure of N. Structural studies have been interpreted to indicate that the group N interacts with a subsite, B2, on the enzyme that comprises Gln69, Asn71 and Glu111. We report studies by site-directed mutagenesis that indicate that Gln69 is not involved in productive interactions with any of the dinucleotide substrates and that Asn71 is an important component of subsite B2 for all dinucleotide substrates tested. Glu111 appears to be functionally involved in catalysis for dinucleotide substrates solely when N is guanosine. |
| Databáze: | OpenAIRE |
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