Subcellular distribution and activation by non-ionic detergents of guanylate cyclase in cerebral cortex of rat.
Non-ionic detergents stimulated particulate guanylate cyclase activity in cerebral cortex of rat 8- to 12-fold while stimulation of soluble enzyme was 1.3- to 2.5-fold. Among various detergents, Lubrol PX was the most effective one. The subcellular distribution of guanylate cyclase activity was examined with or without 0.5% Lubrol PX. Without Lubrol PX two-thirds of the enzyme activity was detected in the soluble fraction. In the presence of Lubrol PX, however, two-thirds of guanylate cyclase activity was recovered in the crude mitochondrial fraction. Further fractionation revealed that most of the particulate guanylate cyclase activity was associated with synaptosomes. The sedimentation characteristic of the particulate guanylate cyclase activity was very close to those of choline acetyltransferase and acetylcholine esterase activities, two synaptosomal enzymes. When the crude mitochondrial fraction was subfractionated after osmotic shock, most of guanylate cyclase activity as assayed in the absence of Lubrol PX was released into the soluble fraction while the rest of the enzyme activity was tightly bound to synaptic membrane fractions. The total guanylate cyclase activity recovered in the synaptosomal soluble fraction was 6 to 7 times higher than that of the starting material. The specific enzyme activity reached more than 1000 pmol per min per mg protein, which was 35-fold higher than that of the starting material. The membrane bound guanylate cyclase activity was markedly stimulated by Lubrol PX. Guanylate cyclase activity in the synaptosomal soluble fraction, in contrast, was suppressed by the addition of Lubrol PX. The observation that most of guanylate cyclase activity was detected in synaptosomes, some of which was tightly bound to the synaptic membrane fraction upon hypoosmotic treatment, is consistent with the concept that cyclic GMP is involved in neural transmission.
Deguchi
T
T
Department of Medical Chemistry, Tokyo Metropolitan Institute for Neurosciences, 2-6 Musashidai, Fuchu-city, Tokyo, Japan 183.
Amano
E
E
Nakane
M
M
D000818:Animals; D002499:Centrifugation, Density Gradient; D002540:Cerebral Cortex / Q000737:chemistry* / Q000201:enzymology*; D003902:Detergents / Q000737:chemistry* / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D006162:Guanylate Cyclase / Q000737:chemistry* / Q000302:isolation & purification; D008297:Male; D008928:Mitochondria / Q000737:chemistry / Q000201:enzymology; D009997:Osmotic Pressure; D000077423:Polidocanol; D011092:Polyethylene Glycols / Q000737:chemistry / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D013347:Subcellular Fractions / Q000737:chemistry / Q000201:enzymology; D013570:Synaptic Membranes / Q000737:chemistry / Q000201:enzymology; D013574:Synaptosomes / Q000737:chemistry / Q000201:enzymology
D003902:Detergents; D000077423:Polidocanol; D011092:Polyethylene Glycols; D006162:Guanylate Cyclase