[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.
Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute at Frederick, MD 21702-1201, United States. zdanov@ncifcrf.gov
A S
AS
Zhdanov
J
J
Phan
A G
AG
Evdokimov
J E
JE
Tropea
R B
RB
Kapust
M
M
Li
A
A
Wlodawer
D S
DS
Waugh
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