[Tobacco etch virus proteinase: crystal structure of the active enzyme and its inactive mutant].
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.
Zhdanov
A S
AS
Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute at Frederick, MD 21702-1201, United States. zdanov@ncifcrf.gov
Phan
J
J
Evdokimov
A G
AG
Tropea
J E
JE
Kapust
R B
RB
Li
M
M
Wlodawer
A
A
Waugh
D S
DS
D001665:Binding Sites; D018360:Crystallography, X-Ray; D010450:Endopeptidases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D017800:Potyvirus / Q000201:enzymology*; D011487:Protein Conformation
D010450:Endopeptidases; C099488:TEV protease