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.
Download full-text PDF |
Source |
|---|
Publication Analysis
Top Keywords
Similar Publications
Structure of the human autophagy factor EPG5 and the molecular basis of its conserved mode of interaction with Atg8-family proteins.
Autophagy
January 2025
Life Sciences Institute, Department of Biochemistry and Molecular Biology, The University of British Columbia, Vancouver, BC, Canada.
The multi-step macroautophagy/autophagy process ends with the cargo-laden autophagosome fusing with the lysosome to deliver the materials to be degraded. The metazoan-specific autophagy factor EPG5 plays a crucial role in this step by enforcing fusion specificity and preventing mistargeting. How EPG5 exerts its critical function and how its deficiency leads to diverse phenotypes of the rare multi-system disorder Vici syndrome are not fully understood.
View Article and Find Full Text PDFSeed-specific expression of AtWRI1 enhanced the yield of cotton seed oil.
Sci Rep
December 2024
School of Biological Sciences, University of the Punjab, Lahore, Pakistan.
The WRINKLED1 (WRI1) transcription factor controls carbon flow in plants through regulating the expression of glycolysis and fatty acid biosynthesis genes. The role of Gossypium hirsutum WRINKLED1 (GhWRI1) in seed-oil accumulation still needs to be explored. Multiple sequence alignment of WRI1 proteins confirmed the presence of two conserved AP2 domains.
View Article and Find Full Text PDFImprovements in large-scale production of tobacco etch virus protease.
Protein Expr Purif
April 2025
Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.
Tobacco-etch-virus (TEV) protease is the workhorse of many laboratories in which protein expression is the linchpin of downstream experiments. TEV protease is remarkable in its sequence specificity as the cleavage sequence rarely appears in higher organisms and its ability to cleave fusion tag proteins from proteins of interest. Herein we report work done on large-scale production of TEV protease using different promotors, media, fusion tags, and expression platforms.
View Article and Find Full Text PDFViral nanoparticles (VNPs) are self-assembled nanometric complexes whose size and shape are similar to those of the virus from which they are derived. VNPs are arousing great attention due to potential biotechnological applications in fields like nanomedicine and nanotechnology because they allow the presentation of polypeptides of choice linked to the virus structural proteins. Starting from tobacco etch virus (TEV), a plant plus-strand RNA virus that belongs to the genus (family ), here we describe the development of recombinant hybrid VNPs in plants able of exposing simultaneously different proteins on their surface.
View Article and Find Full Text PDFProteolysis of host DEAD-box RNA helicase by potyviral proteases activates plant immunity.
New Phytol
February 2025
State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Key Laboratory of Biotechnology in Plant Protection of MARA, Key Laboratory of Green Plant Protection of Zhejiang Province, Institute of Plant Virology, Ningbo University, Ningbo, 315211, China.
Article Synopsis
- The mechanisms by which plant viral proteases interact with host proteins and influence virus-host dynamics are not fully understood, particularly in potyviruses, the largest group of RNA viruses affecting plants.
- Research shows that the protease NIa-Pro from the turnip mosaic virus (TuMV) is crucial for inducing hypersensitive cell death in the plant Nicotiana benthamiana by targeting and degrading the host protein DBP5.
- Additionally, findings reveal that the TuMV-encoded nuclear inclusion b can mitigate NIa-Pro-induced cell death by interacting with DBP5, suggesting complex roles for viral proteases in disrupting host immunity.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!