Structural bases of PAS domain-regulated kinase (PASK) activation in the absence of activation loop phosphorylation

Chintan K. Kikani, Stephen A. Antonysamy, Jeffrey B. Bonanno, Rich Romero, Feiyu Fred Zhang, Marijane Russell, Tarun Gheyi, Miyo Iizuka, Spencer Emtage, J. Michael Sauder, Benjamin E. Turk, Stephen K. Burley, Jared Rutter

Research output: Contribution to journalArticle

19 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)41034-41043
Number of pages10
JournalJournal of Biological Chemistry
Volume285
Issue number52
DOIs
StatePublished - Dec 24 2010

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Phosphorylation
Phosphotransferases
Chemical activation
Protein Kinases
Peptide Library
Mutagenesis
Mammals
Substrates
Site-Directed Mutagenesis
Catalysis
Metabolism
Yeast
Conformations
Catalyst activity
Screening
Yeasts
Crystal structure
Peptides
PAS domain kinases

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Structural bases of PAS domain-regulated kinase (PASK) activation in the absence of activation loop phosphorylation. / Kikani, Chintan K.; Antonysamy, Stephen A.; Bonanno, Jeffrey B.; Romero, Rich; Zhang, Feiyu Fred; Russell, Marijane; Gheyi, Tarun; Iizuka, Miyo; Emtage, Spencer; Sauder, J. Michael; Turk, Benjamin E.; Burley, Stephen K.; Rutter, Jared.

In: Journal of Biological Chemistry, Vol. 285, No. 52, 24.12.2010, p. 41034-41043.

Research output: Contribution to journalArticle

Kikani, CK, Antonysamy, SA, Bonanno, JB, Romero, R, Zhang, FF, Russell, M, Gheyi, T, Iizuka, M, Emtage, S, Sauder, JM, Turk, BE, Burley, SK & Rutter, J 2010, 'Structural bases of PAS domain-regulated kinase (PASK) activation in the absence of activation loop phosphorylation', Journal of Biological Chemistry, vol. 285, no. 52, pp. 41034-41043. https://doi.org/10.1074/jbc.M110.157594
Kikani, Chintan K. ; Antonysamy, Stephen A. ; Bonanno, Jeffrey B. ; Romero, Rich ; Zhang, Feiyu Fred ; Russell, Marijane ; Gheyi, Tarun ; Iizuka, Miyo ; Emtage, Spencer ; Sauder, J. Michael ; Turk, Benjamin E. ; Burley, Stephen K. ; Rutter, Jared. / Structural bases of PAS domain-regulated kinase (PASK) activation in the absence of activation loop phosphorylation. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 52. pp. 41034-41043.
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