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J. Biol. Chem., Vol. 262, Issue 11, 5020-5027, Apr, 1987
KS Wood and LJ Ignarro
The same factors that regulate the activation of purified hepatic soluble
guanylate cyclase by diverse agents possessing distinct requirements for
enzyme activation were found to modulate cyclic GMP formation in intact
viable hepatic cells. A comparison was made between activation of
heme-deficient or heme-reconstituted guanylate cyclase and stimulation of
cyclic GMP formation in mouse hepatic slices that were 95% viable and
showed no active efflux of cyclic GMP. Heme- dependent activators of
guanylate cyclase elicited a greater -fold increase in hepatic cyclic GMP
levels in slices from phenobarbital- pretreated than control mice.
Brilliant cresyl blue and KCN inhibited both enzyme activation and hepatic
cyclic GMP accumulation caused by agents that generate nitric oxide.
Hepatic slices from 3,5- diethoxycarbonyl-1,4-dihydrocollidine-treated
mice, which are known to develop sharp increases in hepatic protoporphyrin
IX/heme concentration ratios, showed elevated resting cyclic GMP levels
whereas phenobarbital pretreatment produced decreased resting cyclic GMP
levels compared to controls. Guanylate cyclase activation by azide required
added catalase, and both enzyme activation and hepatic cyclic GMP formation
were inhibited by aminotriazole. Enzyme activation by glyceryl trinitrate
and NaNO2 required added thiols. Hepatic slices from
acetaminophen-pretreated mice showed marked depletion of sulfhydryls and
decreased cyclic GMP formation in response to these enzyme activators. Both
effects were completely restored by treatment of thiol- depleted mice with
N-acetylcysteine. These observations lend support to the general view that
information gained from studies on the regulatory properties of purified
soluble guanylate cyclase bears a close relationship to studies on
regulatory mechanisms that modulate cyclic GMP formation in intact cells.
Hepatic cyclic GMP formation is regulated by similar factors that modulate activation of purified hepatic soluble guanylate cyclase
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