PMID- 20865444
OWN - NLM
STAT- MEDLINE
DCOM- 20110210
LR  - 20220309
IS  - 1573-6776 (Electronic)
IS  - 0141-5492 (Print)
IS  - 0141-5492 (Linking)
VI  - 32
IP  - 12
DP  - 2010 Dec
TI  - Characterization of the biochemical properties and identification of amino acids 
      forming the catalytic center of 3C-like proteinase of porcine reproductive and 
      respiratory syndrome virus.
PG  - 1905-10
LID - 10.1007/s10529-010-0370-1 [doi]
AB  - The non-structural protein 4 (Nsp4) of porcine reproductive and respiratory 
      syndrome virus (PRRSV) functions as a 3C-like proteinase (3CLpro) and plays a 
      pivotal role in gene expression and replication. We have examined the biochemical 
      properties of PRRSV 3CLpro and identified those amino acid residues involved in 
      its catalytic activity as a prelude to developing anti-PRRSV strategies. The 
      3C-like proteinase (3CLpro) of porcine reproductive and respiratory syndrome 
      virus (PRRSV) was expressed in Escherichia coli and characterized. The optimal 
      temperature and pH for its proteolytic activity were 8 degrees C and 7.5, respectively. 
      Na(+) (1000 mM) and K(+) (500 mM) were not inhibitory to its activity but Cu(2+), 
      Zn(2+), PMSF and EDTA were significantly inhibitory. His(39), Asp(64) and 
      Ser(118) residues were identified to form the catalytic triad of PRRSV 3CLpro by 
      a series of site-directed mutagenesis analysis.
FAU - Xu, Ao-Tian
AU  - Xu AT
AD  - Division of Swine Infectious Diseases, Shanghai Veterinary Research Institute, 
      Chinese Academy of Agricultural Sciences, No. 518, Ziyue Road, Minhang District, 
      Shanghai, 200241, China.
FAU - Zhou, Yan-Jun
AU  - Zhou YJ
FAU - Li, Guo-Xin
AU  - Li GX
FAU - Yu, Hai
AU  - Yu H
FAU - Yan, Li-Ping
AU  - Yan LP
FAU - Tong, Guang-Zhi
AU  - Tong GZ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20100924
PL  - Netherlands
TA  - Biotechnol Lett
JT  - Biotechnology letters
JID - 8008051
RN  - 0 (Amino Acids)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Viral Nonstructural Proteins)
RN  - EC 3.4.22.- (Cysteine Endopeptidases)
SB  - IM
MH  - Amino Acids/*genetics
MH  - *Catalytic Domain
MH  - Cold Temperature
MH  - Cysteine Endopeptidases/chemistry/*genetics/*metabolism
MH  - Enzyme Inhibitors/metabolism
MH  - Enzyme Stability
MH  - Escherichia coli/genetics
MH  - Gene Expression
MH  - Hydrogen-Ion Concentration
MH  - Mutagenesis, Site-Directed
MH  - Porcine respiratory and reproductive syndrome virus/*enzymology/*genetics
MH  - Recombinant Proteins/chemistry/genetics/isolation & purification/metabolism
MH  - Viral Nonstructural Proteins/chemistry/genetics/metabolism
PMC - PMC7088359
EDAT- 2010/09/25 06:00
MHDA- 2011/02/11 06:00
PMCR- 2020/03/23
CRDT- 2010/09/25 06:00
PHST- 2010/05/11 00:00 [received]
PHST- 2010/07/28 00:00 [accepted]
PHST- 2010/09/25 06:00 [entrez]
PHST- 2010/09/25 06:00 [pubmed]
PHST- 2011/02/11 06:00 [medline]
PHST- 2020/03/23 00:00 [pmc-release]
AID - 370 [pii]
AID - 10.1007/s10529-010-0370-1 [doi]
PST - ppublish
SO  - Biotechnol Lett. 2010 Dec;32(12):1905-10. doi: 10.1007/s10529-010-0370-1. Epub 
      2010 Sep 24.