Inducible NO synthase dependent S-nitrosylation and activation of arginase1 contribute to age-related endothelial dysfunction.
Endothelial function is impaired in aging because of a decrease in NO bioavailability. This may be, in part, attributable to increased arginase activity, which reciprocally regulates NO synthase (NOS) by competing for the common substrate, L-arginine. However, the high Km of arginase (>1 mmol/L) compared with NOS (2 to 20 micromol/L) seemingly makes direct competition for substrate unlikely. One of the mechanisms by which NO exerts its effects is by posttranslational modification through S-nitrosylation of protein cysteines. We tested the hypothesis that arginase1 activity is modulated by this mechanism, which serves to alter its substrate affinity, allowing competition with NOS for L-arginine. We demonstrate that arginase1 activity is altered by S-nitrosylation, both in vitro and ex vivo. Furthermore, using site-directed mutagenesis we demonstrate that 2 cysteine residues (C168 and C303) are able to undergo nitrosylation. S-Nitrosylation of C303 stabilizes the arginase1 trimer and reduces its Km value 6-fold. Finally, arginase1 nitrosylation is increased (and thus its Km decreased) in blood vessels from aging rats, likely contributing to impaired NO bioavailability and endothelial dysfunction. This is mediated by inducible NOS, which is expressed in the aging endothelium. These findings suggest that S-nitrosylated arginase1 can compete with NOS for L-arginine and contribute to endothelial dysfunction in the aging cardiovascular system.
Santhanam
Lakshmi
L
Johns Hopkins University School of Medicine, 720 Rutland Ave, Traylor 621, Baltimore MD 21209, USA.
Lim
Hyun Kyo
HK
Lim
Hyun Kyoung
HK
Miriel
Victor
V
Brown
Tashalee
T
Patel
Meet
M
Balanson
Sarit
S
Ryoo
Sungwoo
S
Anderson
Mirinda
M
Irani
Kaikobad
K
Khanday
Firdous
F
Di Costanzo
Luigi
L
Nyhan
Daniel
D
Hare
Joshua M
JM
Christianson
David W
DW
Rivers
Richard
R
Shoukas
Artin
A
Berkowitz
Dan E
DE
D000367:Age Factors; D000375:Aging / Q000378:metabolism* / Q000473:pathology; D000818:Animals; D001119:Arginase / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D004730:Endothelium, Vascular / Q000201:enzymology* / Q000473:pathology; D004789:Enzyme Activation / Q000235:genetics; D006801:Humans; D008297:Male; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D009584:Nitrogen / Q000378:metabolism; D016716:PC12 Cells; D051381:Rats; D017208:Rats, Wistar; D014652:Vascular Diseases / Q000201:enzymology* / Q000235:genetics / Q000473:pathology
D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; D001119:Arginase; D009584:Nitrogen