scholarly journals CPDadh: A new peptidase family homologous to the cysteine protease domain in bacterial MARTX toxins

2009 ◽  
pp. NA-NA
Author(s):  
Jimin Pei ◽  
Patrick J. Lupardus ◽  
K. Christopher Garcia ◽  
Nick V. Grishin
Keyword(s):  
Cysteine Protease ◽  
Protease Domain ◽  
Cysteine Protease Domain

scholarly journals The Porcine Reproductive and Respiratory Syndrome Virus nsp2 Cysteine Protease Domain Possesses both trans- and cis-Cleavage Activities

2009 ◽  
Vol 83 (18) ◽  
pp. 9449-9463 ◽  
Author(s):  
Jun Han ◽  
Mark S. Rutherford ◽  
Kay S. Faaberg
Keyword(s):  
Cysteine Protease ◽  
Reverse Genetics ◽  
Nonstructural Protein ◽  
Point Mutations ◽  
Respiratory Syndrome Virus ◽  
Protease Domain ◽  
Nonstructural Protein 2 ◽  
Cleavage Activity ◽  
Cysteine Protease Domain

ABSTRACT The N terminus of the replicase nonstructural protein 2 (nsp2) of porcine reproductive and respiratory syndrome virus (PRRSV) contains a putative cysteine protease domain (PL2). Previously, we demonstrated that deletion of either the PL2 core domain (amino acids [aa] 47 to 180) or the immediate downstream region (aa 181 to 323) is lethal to the virus. In this study, the PL2 domain was found to encode an active enzyme that mediates efficient processing of nsp2-3 in CHO cells. The PL2 protease possessed both trans- and cis-cleavage activities, which were distinguished by individual point mutations in the protease domain. The minimal size required to maintain these two enzymatic activities included nsp2 aa 47 to 240 (Tyr47 to Cys240) and aa 47 to 323 (Tyr47 to Leu323), respectively. Introduction of targeted amino acid mutations in the protease domain confirmed the importance of the putative Cys55- His124 catalytic motif for nsp2/3 proteolysis in vitro, as were three additional conserved cysteine residues (Cys111, Cys142, and Cys147). The conserved aspartic acids (e.g., Asp89) were essential for the PL2 protease trans-cleavage activity. Reverse genetics revealed that the PL2 trans-cleavage activity played an important role in the PRRSV replication cycle in that mutations that impaired the PL2 protease trans function, but not the cis activity, were detrimental to viral viability. Lastly, the potential nsp2/3 cleavage site was probed. Mutations with the largest impact on in vitro cleavage were at or near the G1196|G1197 dipeptide.

scholarly journals Autoprocessing of the Vibrio cholerae RTX toxin by the cysteine protease domain

2007 ◽  
Vol 26 (10) ◽  
pp. 2552-2561 ◽  
Author(s):  
Kerri-Lynn Sheahan ◽  
Christina L Cordero ◽  
Karla J Fullner Satchell
Keyword(s):  
Vibrio Cholerae ◽  
Cysteine Protease ◽  
Protease Domain ◽  
Rtx Toxin ◽  
Cysteine Protease Domain

scholarly journals Small Molecule-Induced Allosteric Activation of the Vibrio cholerae RTX Cysteine Protease Domain

Science ◽  
2008 ◽  
Vol 322 (5899) ◽  
pp. 265-268 ◽  
Author(s):  
P. J. Lupardus ◽  
A. Shen ◽  
M. Bogyo ◽  
K. C. Garcia
Keyword(s):  
Vibrio Cholerae ◽  
Small Molecule ◽  
Cysteine Protease ◽  
Protease Domain ◽  
Allosteric Activation ◽  
Cysteine Protease Domain

scholarly journals The Cysteine Protease Domain of Porcine Reproductive and Respiratory Syndrome Virus Nonstructural Protein 2 Possesses Deubiquitinating and Interferon Antagonism Functions

2010 ◽  
Vol 84 (15) ◽  
pp. 7832-7846 ◽  
Author(s):  
Zhi Sun ◽  
Zhenhai Chen ◽  
Steven R. Lawson ◽  
Ying Fang
Keyword(s):  
Cysteine Protease ◽  
Nonstructural Protein ◽  
Cell Epitope ◽  
Wild Type Virus ◽  
Type I ◽  
Protease Domain ◽  
Live Virus ◽  
Recombinant Viruses ◽  
Nonstructural Protein 2 ◽  
Cysteine Protease Domain

ABSTRACT Porcine reproductive and respiratory syndrome (PRRS) virus nonstructural protein 2 (nsp2) contains a cysteine protease domain at its N terminus, which belongs to the ovarian tumor (OTU) protease family. In this study, we demonstrated that the PRRSV nsp2 OTU domain antagonizes the type I interferon induction by interfering with the NF-κB signaling pathway. Further analysis revealed that the nsp2 OTU domain possesses ubiquitin-deconjugating activity. This domain has the ability to inhibit NF-κB activation by interfering with the polyubiquitination process of IκBα, which subsequently prevents IκBα degradation. To determine whether the nsp2 protein antagonist function can be ablated from the virus, we introduced point mutations into the OTU domain region by use of reverse genetics. The D458A, S462A, and D465A mutations targeting on a B-cell epitope in the OTU domain region generated the viable recombinant viruses, and the S462A and D465A mutants were attenuated for growth in cell culture. The OTU domain mutants were examined to determine whether mutations in the nsp2 OTU domain region altered virus ability to inhibit NF-κB activation. The result showed that certain mutations lethal to virus replication impaired the ability of nsp2 to inhibit NF-κB activation but that the viable recombinant viruses, vSD-S462A and vSD-D465A, were unable to inhibit NF-κB activation as effectively as the wild-type virus. This study represents a fundamental step in elucidating the role of nsp2 in PRRS pathogenesis and provides an important insight in future modified live-virus vaccine development.

scholarly journals Vibrio Type III Effector VPA1380 Is Related to the Cysteine Protease Domain of Large Bacterial Toxins

PLoS ONE ◽  
2014 ◽  
Vol 9 (8) ◽  
pp. e104387 ◽  
Author(s):  
Thomas Calder ◽  
Lisa N. Kinch ◽  
Jessie Fernandez ◽  
Dor Salomon ◽  
Nick V. Grishin ◽  
...  
Keyword(s):  
Cysteine Protease ◽  
Bacterial Toxins ◽  
Protease Domain ◽  
Type Iii Effector ◽  
Type Iii ◽  
Cysteine Protease Domain

scholarly journals Defining an allosteric circuit in the cysteine protease domain of Clostridium difficile toxins

2011 ◽  
Vol 18 (3) ◽  
pp. 364-371 ◽  
Author(s):  
Aimee Shen ◽  
Patrick J Lupardus ◽  
Malte M Gersch ◽  
Aaron W Puri ◽  
Victoria E Albrow ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Cysteine Protease ◽  
Protease Domain ◽  
Cysteine Protease Domain
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