Site-directed mutagenesis of the glycine-rich loop of death associated protein kinase (DAPK) identifies it as a key structure for catalytic activity

Laurie K. McNamara, Joseph S. Brunzelle, James P. Schavocky, D. Martin Watterson, Valerie Grum-Tokars*

*Corresponding author for this work

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Death associated protein kinase (DAPK) is a calmodulin (CaM)-regulated protein kinase that is a therapeutic target for central nervous system (CNS) disorders. We report here the results of studies that test the hypothesis of McNamara et al. (2009) that conformational selection in DAPK's glycine-rich region is key for catalytic activity. The hypothesis was tested by site-directed mutagenesis of glutamine-23 (Q23) in the middle of this loop. The glycine-rich loop exhibits localized differences in structure among DAPK conformations that correlate with different stages of the catalytic cycle. Changing the Q23 to a Valine (V23), found at the corresponding position in another CaM regulated protein kinase, results in a reduced catalytic efficiency. High resolution X-ray crystal structures of various conformations of the Q23V mutant DAPK and their superimposition with the corresponding conformations from wild type catalytic domain reveal localized changes in the glycine-rich region. The effect of the mutation on DAPK catalytic activity and the finding of only localized changes in the DAPK structure provide experimental evidence implicating conformational selection in this domain with activity. This article is part of a Special Issue entitled: 11th European Symposium on Calcium.

Original languageEnglish (US)
Pages (from-to)1068-1073
Number of pages6
JournalBiochimica et Biophysica Acta - Molecular Cell Research
Volume1813
Issue number5
DOIs
StatePublished - May 1 2011

Keywords

  • Calmodulin
  • Central nervous system
  • Death associated protein kinase
  • Glycine-rich region
  • Neurodegeneration
  • Phosphorylation
  • Protein kinase inhibitor

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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