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 language | English (US) |
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Pages (from-to) | 1068-1073 |
Number of pages | 6 |
Journal | Biochimica et Biophysica Acta - Molecular Cell Research |
Volume | 1813 |
Issue number | 5 |
DOIs | |
State | Published - May 2011 |
Funding
This work was supported in part by NIH awards NS056051 and AG031311 (DMW).
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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Crystal Structure of DAPK1 catalytic domain in complex with the hinge binding fragment phthalazine
McNamara, L. K. (Contributor), Brunzelle, J. S. (Contributor), Schavocky, J. P. (Contributor), Watterson, D. M. (Contributor) & Grum-Tokars, V. (Contributor), Protein Data Bank (PDB), May 6 2015
DOI: 10.2210/pdb4YO4/pdb, https://www.wwpdb.org/pdb?id=pdb_00004yo4
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