Identification of nitric oxide synthase in Staphylococcus aureus.
The presence of nitric oxide synthase (NOS) in Staphylococcus aureus ATCC6538P was established by western blot analysis. The identity of citrulline formed from L-arginine by the NOS was confirmed by both TLC and HPLC and the other product, NO, by directly measuring the production of nitrogen oxides (NOx) in the reaction mixture. The activity was inhibited by typical NOS inhibitors such as N-nitro-L-arginine methylester and NG,NG-dimethyl-L-arginine with the IC50 of 4.9 x 10(-4) and 2.5 x 10(-4) M, respectively. The NOS activity was maximum at pH 6.5 and at 47.5 degrees C. These results indicate that the NOS of S. aureus ATCC6538P is an isoform distinct from mammalian NOS.
Choi
W S
WS
College of Pharmacy, Sung Kyun Kwan University, Suwon City, Kyunggi-Do, Korea.
Chang
M S
MS
Han
J W
JW
Hong
S Y
SY
Lee
H W
HW
D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D002956:Citrulline / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D009249:NADP / Q000378:metabolism; D009569:Nitric Oxide / Q000032:analysis; D019001:Nitric Oxide Synthase / Q000302:isolation & purification / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000201:enzymology*; D013816:Thermodynamics
D002956:Citrulline; D009569:Nitric Oxide; D009249:NADP; D019001:Nitric Oxide Synthase