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Direct interaction between the reductase domain of endothelial nitric oxide synthase and the ryanodine receptor
Author(s) -
Martínez-Moreno Mónica,
Álvarez-Barrientos Alberto,
Roncal Fernando,
Albar Juan Pablo,
Gavilanes Francisco,
Lamas Santiago,
Rodríguez-Crespo Ignacio
Publication year - 2005
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2005.04.078
Subject(s) - ryanodine receptor , enos , nitric oxide , nitric oxide synthase , immunoprecipitation , chemistry , nitric oxide synthase type iii , microbiology and biotechnology , receptor , biochemistry , biology , enzyme , organic chemistry , gene
We have performed the recombinant expression and purification of the reductase domain of endothelial nitric oxide synthase (eNOS) and used it as a bait in search for interacting proteins present in endothelial cells. Using mass spectrometry of the bound proteins run in a PAGE–SDS gel, we were able to identify the ryanodine receptor (RyR) as a novel eNOS‐binding partner. This interaction was confirmed through immunoprecipitation of both RyR and eNOS from endothelial cells and cardiac myocytes. Immunofluorescence data indicated that a subpopulation of eNOS associates with RyR in perinuclear regions of the cell, where eNOS might be responsible for the known nitrosylation of RyR.