PMID- 17913645
OWN - NLM
STAT- MEDLINE
DCOM- 20080128
LR  - 20091119
IS  - 0076-6879 (Print)
IS  - 0076-6879 (Linking)
VI  - 430
DP  - 2007
TI  - Biophysical and biochemical investigations of dsRNA-activated kinase PKR.
PG  - 373-96
AB  - Protein kinase RNA-activated (PKR) is a serine/threonine kinase that contains an 
      N-terminal RNA-binding domain (dsRNA) and a C-terminal kinase domain. On binding 
      viral dsRNA molecules, PKR can become activated and phosphorylate cellular 
      targets, such as eukaryotic translation initiation factor 2alpha (eIF-2alpha). 
      Phosphorylation of eIF-2alpha results in attenuation of protein translation 
      initiation. Therefore, PKR plays an integral role in the antiviral response to 
      cellular infection. Here we provide a methodological framework for probing PKR 
      function by use of assays for phosphorylation, RNA-protein stability, PKR 
      dimerization, and in vitro translation. These methods are complemented by nuclear 
      magnetic resonance approaches for probing structural features of PKR activation. 
      Considerations required for both PKR and dsRNA sample preparation are also 
      discussed.
FAU - McKenna, Sean A
AU  - McKenna SA
AD  - Department of Structural Biology, Stanford University School of Medicine, 
      Stanford, California, USA.
FAU - Lindhout, Darrin A
AU  - Lindhout DA
FAU - Shimoike, Takashi
AU  - Shimoike T
FAU - Puglisi, Joseph D
AU  - Puglisi JD
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Methods Enzymol
JT  - Methods in enzymology
JID - 0212271
RN  - 0 (RNA, Double-Stranded)
RN  - 63231-63-0 (RNA)
RN  - EC 2.7.11.1 (eIF-2 Kinase)
SB  - IM
MH  - Humans
MH  - Nuclear Magnetic Resonance, Biomolecular
MH  - Phosphorylation
MH  - Protein Biosynthesis
MH  - *RNA/chemistry/metabolism
MH  - RNA Stability
MH  - RNA, Double-Stranded/metabolism
MH  - *eIF-2 Kinase/chemistry/genetics/metabolism
EDAT- 2007/10/05 09:00
MHDA- 2008/01/29 09:00
CRDT- 2007/10/05 09:00
PHST- 2007/10/05 09:00 [pubmed]
PHST- 2008/01/29 09:00 [medline]
PHST- 2007/10/05 09:00 [entrez]
AID - S0076-6879(07)30014-1 [pii]
AID - 10.1016/S0076-6879(07)30014-1 [doi]
PST - ppublish
SO  - Methods Enzymol. 2007;430:373-96. doi: 10.1016/S0076-6879(07)30014-1.