Journal
JOURNAL OF BIOLOGICAL CHEMISTRY
Volume 278, Issue 45, Pages 44719-44726Publisher
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.M302836200
Keywords
-
Categories
Funding
- NHLBI NIH HHS [HL61371, HL64793, R01 HL57665, HL71214, HL68769, HL 61417] Funding Source: Medline
Ask authors/readers for more resources
There is evidence that endothelial nitric-oxide synthase ( eNOS) is regulated by reciprocal dephosphorylation of Thr(497) and phosphorylation of Ser(1179). To examine the interrelationship between these sites, cells were transfected with wild-type (WT), T497A, T497D, S1179D, and T497A/S1179D eNOS and activity, NO release and eNOS localization were assessed. Although eNOS T497A, S1179D and T497A/S1179D eNOS had greater enzymatic activity than did WT eNOS in lysates, basal production of NO from cells was markedly reduced in cells transfected with T497A and T497A/S1179D eNOS but augmented in cells transfected with S1179D eNOS. Stimulating cells with ATP or ionophore normalized the loss of function seen with T497A and T497A/S1179D eNOS to levels observed with WT and S1179D eNOS, respectively. Despite these functional differences, the localization of eNOS mutants were similar to WT. Because both T497A and T497A/S1179D eNOS exhibited higher enzyme activity but reduced production of NO, we examined whether these mutations were uncoupling NO synthesis. T497A and T497A/S1179D eNOS generated 2 - 3 times more superoxide anion than WT eNOS, and both basal and stimulated interactions of T497A/S1179D eNOS with hsp90 were reduced in co- immunoprecipitation experiments. Thus, the phosphorylation/dephosphorylation of Thr497 may be an intrinsic switch mechanism that determines whether eNOS generates NO versus superoxide in cells.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available