[Tobacco etch virus proteinase: crystal structure of the active enzyme and its inactive mutant].

Abstract

Tobacco Etch Virus Protease (TEV protease) is widely used as a tool for separation of recombinant target proteins from their fusion partners. The crystal structures of two mutants of TEV protease, active autolysis-resistant mutant TEV-S219D in complex with the proteolysis product, and inactive mutant TEV-C151A in complex with a substrate, have been determined at 1.8 and 2.2 A resolution, respectively. The active sites of both mutants, including their oxyanion holes, have identical structures. The C-terminal residues 217-221 of the enzyme are involved in formation of the binding pockets S3-S6. This indicates that the autolysis of the peptide bond Met218-Ser219 exerts a strong effect on the fine-tuning of the substrate in the enzyme active site, which results in considerable decrease in the enzymatic activity.

• Authors: A S Zhdanov, J Phan, A G Evdokimov, J E Tropea, R B Kapust, M Li, A Wlodawer, D S Waugh
• Journal: Bioorganicheskaia khimiia (Bioorg Khim) 2003 Sep-Oct Vol. 29 Issue 5 Pg. 457-60 ISSN: 0132-3423 [Print] Russia (Federation)
• Vernacular Title: Proteinaza virusa gravirovki tabaka: kristallicheskaia struktura aktivnogo fermenta i ego neaktivnogo mutanta.
• PMID: 14601399 (Publication Type: English Abstract, Journal Article)

Chemical References

• Endopeptidases
• TEV protease

Topics

• Binding Sites
• Crystallography, X-Ray
• Endopeptidases (chemistry, genetics, metabolism)
• Mutation
• Potyvirus (enzymology)
• Protein Conformation