The phosphorylation motif at serine 225 governs the localization and function of sphingosine kinase 1 in resistance arteries
(2009) In Arteriosclerosis, Thrombosis, and Vascular Biology 29(11). p.1916-1922- 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... (More)
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.
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- author
- Lidington, Darcy ; Peter, Bernhard Friedrich ; Meissner, Anja LU ; Kroetsch, Jeffrey T. ; Pitson, Stuart M. ; Pohl, Ulrich and Bolz, Steffen Sebastian
- publishing date
- 2009-11
- type
- Contribution to journal
- publication status
- published
- keywords
- ERK1/2, Myogenic vasoconstriction, Signal transduction, Sphingosine-1-phosphate, Transfection
- in
- Arteriosclerosis, Thrombosis, and Vascular Biology
- volume
- 29
- issue
- 11
- pages
- 7 pages
- publisher
- Lippincott Williams & Wilkins
- external identifiers
-
- scopus:73949131765
- pmid:19729605
- ISSN
- 1079-5642
- DOI
- 10.1161/ATVBAHA.109.194803
- language
- English
- LU publication?
- no
- id
- 49f8a710-7b34-4473-853b-3749c30a0e6b
- date added to LUP
- 2017-05-23 22:25:48
- date last changed
- 2024-01-13 21:37:09
@article{49f8a710-7b34-4473-853b-3749c30a0e6b, abstract = {{<p>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 Sk1<sup>s225A</sup> reduced resting and myogenic tone, resting Ca<sup>2+</sup>, pressure-induced Ca<sup>2+</sup> elevations, and Ca <sup>2+</sup> sensitivity. The lack of function of the Sk1<sup>s225A</sup> 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, Ca<sup>2+</sup> sensitivity and myogenic tone were attenuated, whereas Ca <sup>2+</sup> handling was normalized/enhanced. These discrete effects are consistent with cell surface receptor-mediated effects (Ca<sup>2+</sup> sensitivity) and intracellular effects of S1P (Ca<sup>2+</sup> handling). Accordingly, S1P2 receptor inhibition (1μmol/L JTE013) attenuated myogenic tone without effect on Ca<sup>2+</sup>. 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.</p>}}, author = {{Lidington, Darcy and Peter, Bernhard Friedrich and Meissner, Anja and Kroetsch, Jeffrey T. and Pitson, Stuart M. and Pohl, Ulrich and Bolz, Steffen Sebastian}}, issn = {{1079-5642}}, keywords = {{ERK1/2; Myogenic vasoconstriction; Signal transduction; Sphingosine-1-phosphate; Transfection}}, language = {{eng}}, number = {{11}}, pages = {{1916--1922}}, publisher = {{Lippincott Williams & Wilkins}}, series = {{Arteriosclerosis, Thrombosis, and Vascular Biology}}, title = {{The phosphorylation motif at serine 225 governs the localization and function of sphingosine kinase 1 in resistance arteries}}, url = {{http://dx.doi.org/10.1161/ATVBAHA.109.194803}}, doi = {{10.1161/ATVBAHA.109.194803}}, volume = {{29}}, year = {{2009}}, }