Abstract
Publisher Summary This chapter discusses about the organic nitrate reductase. Organic nitrate reductase catalyzes the reduction of various nitrate esters by glutathione. Inorganic nitrite is liberated and the assay procedure is based on its rate of formation. For nitrite analysis a mercuric chloride filtrate is prepared. The assay method is applicable to crude tissue preparations. Such preparations frequently form inorganic nitrite from nitroglycerine without addition of GSH, but after dialysis GSH becomes a necessary component of the assay system. The steps described in the purification procedure are preparation of crude extract, first ammonium sulfate fractionation, second ammonium sulfate fractionation, ethanol fractionation, third ammonium sulfate fractionation, and adsorption on calcium phosphate gel. The purified enzyme has been found to be active with both nitroglycerin and erythritol tetranitrate. For the enzyme-catalyzed reaction GSH cannot be replaced by cysteine. However, there is a slow spontaneous reaction between both GSH and cysteine, on the one hand, and the nitrate esters. This occurs more rapidly in alkali. Balance studies show that 2 micromoles of GSH are oxidized for every micromole of nitrite formed.
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