Mapping of the Auto-inhibitory Interactions of Protein Kinase R by Nuclear Magnetic Resonance

Vladimir Gelev, Huseyin Aktas, Assen Marintchev, Takuhiro Ito, Dominique Frueh, Michael Hemond, David Rovnyak, Mirijam Debus, Sven Hyberts, Anny Usheva, Jose Halperin, Gerhard Wagner

Research output: Contribution to journalArticlepeer-review


The dsRNA-dependent protein kinase (PKR) is a key mediator of the anti-viral and anti-proliferative effects of interferon. Unphosphorylated PKR is characterized by inhibitory interactions between the kinase and RNA binding domains (RBDs), but the structural details of the latent state and its unraveling during activation are not well understood. To study PKR regulation by NMR we assigned a large portion of the backbone resonances of the catalytically inactive K296R kinase domain, and performed 15N-heteronuclear single quantum coherence (HSQC) titrations of this kinase domain with the RBDs. Chemical shift perturbations in the kinase indicate that RBD2 binds to the substrate eIF2α docking site in the kinase C-lobe. Consistent with these results, a mutation in the eIF2α docking site, F495A, displays weaker interactions with the RBD. The full-length RBD1 + 2 binds more strongly to the kinase domain than RBD2 alone. The observed chemical shift changes extend from the eIF2α binding site into the kinase N-lobe and inside the active site, consistent with weak interactions between the N-terminal part of the RBD and the kinase.

Original languageEnglish (US)
Pages (from-to)352-363
Number of pages12
JournalJournal of molecular biology
Issue number3
StatePublished - Dec 1 2006
Externally publishedYes


  • NMR: kinase
  • PKR
  • RNA-binding domain
  • elF2α

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology


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