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Structural Bases of PAS Domain-regulated Kinase (PASK) Activation in the Absence of Activation Loop Phosphorylation
- Source :
- Journal of Biological Chemistry. 285:41034-41043
- Publication Year :
- 2010
- Publisher :
- Elsevier BV, 2010.
-
Abstract
- Per-Arnt-Sim (PAS) domain-containing protein kinase (PASK) is an evolutionary conserved protein kinase that coordinates cellular metabolism with metabolic demand in yeast and mammals. The molecular mechanisms underlying PASK regulation, however, remain unknown. Herein, we describe a crystal structure of the kinase domain of human PASK, which provides insights into the regulatory mechanisms governing catalysis. We show that the kinase domain adopts an active conformation and has catalytic activity in vivo and in vitro in the absence of activation loop phosphorylation. Using site-directed mutagenesis and structural comparison with active and inactive kinases, we identified several key structural features in PASK that enable activation loop phosphorylation-independent activity. Finally, we used combinatorial peptide library screening to determine that PASK prefers basic residues at the P-3 and P-5 positions in substrate peptides. Our results describe the key features of the PASK structure and how those features are important for PASK activity and substrate selection.
- Subjects :
- Protein-Serine-Threonine Kinases
Kinase
Cell Biology
Protein Serine-Threonine Kinases
Biology
Biochemistry
Protein Structure, Secondary
Protein Structure, Tertiary
Enzyme Activation
Structure-Activity Relationship
Enzyme activator
Protein kinase domain
PAS domain
Protein Structure and Folding
Mutagenesis, Site-Directed
Humans
Phosphorylation
Protein phosphorylation
Protein kinase A
Molecular Biology
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 285
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Chemistry
- Accession number :
- edsair.doi.dedup.....35975780d2184c10f5d049993cde0da5
- Full Text :
- https://doi.org/10.1074/jbc.m110.157594