B. Mcdonald et al., GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE IS REQUIRED FOR THE TRANSPORT OF NITRIC-OXIDE IN PLATELETS, Proceedings of the National Academy of Sciences of the United Statesof America, 90(23), 1993, pp. 11122-11126
Nitric oxide (NO) or NO-generating compounds like sodium nitroprusside
(SNP) increase cellular levels of cGMP and produce S-nitrosylation of
glyceraldehyde-3-phosphate dehydrogenase [GAPDH; D-glyceraldehyde-3-p
hosphate: NAD+ oxidoreductase (phosphorylating), EC 1.2.1.12]. In,sear
ch of a reagent that could discriminate between these two effects, we
used the sesquiterpene antibiotic koningic acid, which binds to GAPDH
at the Cys-149 of the active site. Koningic acid inhibited basal and s
odium nitroprusside-stimulated NAD-dependent covalent modification of
purified rabbit muscle GAPDH in a dose-dependent manner. Furthermore,
we tested the effect of koningic acid on human platelets. Approximatel
y 90% of GAPDH is present in the cytosol of human platelets, and the e
xposure of platelet cytosol to koningic acid inhibited GAPDH activity,
while the soluble guanylyl cyclase (basal and sodium nitroprusside-st
imulated) activity remained unaltered. Pretreatment of intact platelet
s with koningic acid slowed the rate of aggregation induced by a subma
ximal concentration of thrombin. In addition, the antibiotic also inhi
bited the cGMP increases triggered by SNP, S-nitroso-N-acetylpenicilla
mine (SNAP), and 3-morpholinosydnomidine (SIN-1) but failed to prevent
an increase in cGMP caused by nitrosylated albumin. Under the same co
nditions, koningic acid also inhibited basal and SNP- SNAP-, and SIN-1
-stimulated NAD-dependent modification of GAPDH and its enzymatic acti
vity. These results suggest that the mechanism of delivery of NO from
SNP, SNAP, and SIN-1 to platelets may require the active form of GAPDH
. When NO is delivered by nitrosylated albumin, active GAPDH was not n
ecessary.