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J. Biochem, 1990, Vol. 107, No. 5 694-698
© 1990 Japanese Biochemical Society

research-article

Subcellular Distribution of GTP-Binding Proteins, Go and G12, in Rat Cerebral Cortex1

Tomiko Asano, Masato Nagahama and Kanefusa Kato

Institute for Developmental Research, Aichi Prefectural Colony Kasugai, Aichi 480–03

The localization of GTP-binding protein (G-protein) subunits, Go, G12a and ß, in subcellular fractions of rat cerebral cortex was determined by means of immunoassays specific for the respective subunits. High concentrations of all three subunits were observed in both crude mitochondrial and microsomal fractions. Muscarinic cholinergic receptors were also densely localized in these fractions. Then the crude mitochondrial and microsomal fractions were subfractionated by sucrose density gradient centrifugation. Each fraction obtained was evaluated morphologically by electron microscopy and biochemically by determination of membrane markers. The crude mitochondrial fraction was subfractionated into myelin, synaptic plasma membrane, and mitochondrial fractions. All the G-protein subunits examined and muscarinic receptors were exclusively localized in the synaptic plasma membrane fraction. Among the submicrosomal fractions, the heavy smooth-surfaced microsomal fraction showed the highest concentrations of all G-protein subunits and receptors, while the rough-surfaced microsomal fraction contained low amounts of them. The heavy smooth-surfaced microsomal fraction also contained high specific activity of (Na+-K+)-ATPase, a marker of the plasma membrane. These results indicated that the Go, Ga2 and ß subunits are mainly localized in the plasma membrane in the brain.

1 This work was supported in part by a Grant-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Science and Culture of Japan, and a research grant from the Ishida Foundation. Abbreviations: G-proteins, GTP-binding protein; Gh the GTP-binding protein whose function is in part the mediation of inhibition of adenylate cyclase; Go, a related GTP-binding protein of unknown function; QNB, quinuclidinyl benzilate.


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