S-Glutathiolation by peroxynitrite activates SERCA during arterial relaxation by nitric oxide.
Nitric oxide (NO) physiologically stimulates the sarco/endoplasmic reticulum calcium (Ca(2+)) ATPase (SERCA) to decrease intracellular Ca(2+) concentration and relax cardiac, skeletal and vascular smooth muscle. Here, we show that NO-derived peroxynitrite (ONOO(-)) directly increases SERCA activity by S-glutathiolation and that this modification of SERCA is blocked by irreversible oxidation of the relevant cysteine thiols during atherosclerosis. Purified SERCA was S-glutathiolated by ONOO(-) and the increase in Ca(2+)-uptake activity of SERCA reconstituted in phospholipid vesicles required the presence of glutathione. Mutation of the SERCA-reactive Cys674 to serine abolished these effects. Because superoxide scavengers decreased S-glutathiolation of SERCA and arterial relaxation by NO, ONOO(-) is implicated as the intracellular mediator. NO-dependent relaxation as well as S-glutathiolation and activation of SERCA were decreased by atherosclerosis and Cys674 was found to be oxidized to sulfonic acid. Thus, irreversible oxidation of key thiol(s) in disease impairs NO-induced relaxation by preventing reversible S-glutathiolation and activation of SERCA by NO/ONOO(-).
Vascular and Myocardial Biology Units, Whitaker Cardiovascular Institute, Boston University Medical Center, X707, 650 Albany Street, Boston, Massachusetts 02118-2393, USA. Tadachi@sc.itc.keio.ac.jp
Takeshi
T
Adachi
Robert M
RM
Weisbrod
David R
DR
Pimentel
Jia
J
Ying
Victor S
VS
Sharov
Christian
C
Schöneich
Richard A
RA
Cohen
D000818:Animals; D001161:Arteriosclerosis / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers; D005978:Glutathione / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D051379:Mice; D008822:Mice, Transgenic; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D016296:Mutagenesis; D009154:Mutation / Q000235:genetics; D009569:Nitric Oxide / Q000378:metabolism*; D030421:Peroxynitrous Acid / Q000378:metabolism*; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases
D017931:DNA Primers; D030421:Peroxynitrous Acid; D009569:Nitric Oxide; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D000252:Calcium-Transporting ATPases; D005978:Glutathione; D002118:Calcium