The phosphorylation motif at serine 225 governs the localization and function of sphingosine kinase 1 in resistance arteries

Research output: Contribution to journalArticle

Abstract

OBJECTIVE-: The purpose of this study was to characterize a phosphorylation motif at serine 225 as a molecular switch that regulates the pressure-dependent activation of sphingosine kinase 1 (Sk1) in resistance artery smooth muscle cells. METHODS AND RESULTS-: In isolated hamster gracilis muscle resistance arteries, pressure-dependent activation/translocation of Sk1 by ERK1/2 was critically dependent on its serine 225 phosphorylation site. Specifically, expression of Sk1s225A reduced resting and myogenic tone, resting Ca2+, pressure-induced Ca2+ elevations, and Ca 2+ sensitivity. The lack of function of the Sk1s225A mutant could not be entirely overcome by forced localization to the plasma membrane via a myristoylation/palmitylation motif; the membrane anchor also significantly inhibited the function of the wild-type Sk1 enzyme. In both cases, Ca2+ sensitivity and myogenic tone were attenuated, whereas Ca 2+ handling was normalized/enhanced. These discrete effects are consistent with cell surface receptor-mediated effects (Ca2+ sensitivity) and intracellular effects of S1P (Ca2+ handling). Accordingly, S1P2 receptor inhibition (1μmol/L JTE013) attenuated myogenic tone without effect on Ca2+. CONCLUSIONS-: Translocation and precise subcellular positioning of Sk1 is essential for full Sk1 function; and two distinct S1P pools, proposed to be intra-and extracellular, contribute to the maintenance of vascular tone.

Details

Authors
  • Darcy Lidington
  • Bernhard Friedrich Peter
  • Anja Meissner
  • Jeffrey T. Kroetsch
  • Stuart M. Pitson
  • Ulrich Pohl
  • Steffen Sebastian Bolz
External organisations
  • World Orthopaedic Concern
  • Institute of Medical and Veterinary Science Australia
  • University of Toronto
Research areas and keywords

Keywords

  • ERK1/2, Myogenic vasoconstriction, Signal transduction, Sphingosine-1-phosphate, Transfection
Original languageEnglish
Pages (from-to)1916-1922
Number of pages7
JournalArteriosclerosis, Thrombosis, and Vascular Biology
Volume29
Issue number11
Publication statusPublished - 2009 Nov
Publication categoryResearch
Peer-reviewedYes
Externally publishedYes