毒蕈碱型乙酰胆碱受体拮抗剂对大鼠肾钠氢交换体的抑制作用及标记

Inhibition and labeling of the rat renal Na+/H+-exchanger by an antagonist of muscarinic acetylcholine receptors.

作者信息

Friedrich T, Burckhardt G

机构信息

Department of Medical Chemistry, Kyoto University Faculty of Medicine, Japan.

出版信息

Biochem Biophys Res Commun. 1988 Dec 30;157(3):921-9. doi: 10.1016/s0006-291x(88)80962-8.

DOI:10.1016/s0006-291x(88)80962-8

PMID:2849947

Abstract

A covalently binding label for muscarinic acetylcholine receptors, propylbenzilylcholine mustard (PrBCM), irreversibly inhibits the Na+/H+ exchanger in rat renal brush-border membrane vesicles. Substrates of the antiporter, Na+ and Li+, as well as inhibitors, amiloride, 5-(N-ethyl-N-isopropyl)amiloride (EIPA) and propranolol, protect the antiporter from inactivation by PrBCM. With [3H]PrBCM a band with an app. Mr of 65 kDa is predominantly labeled. Amiloride protects this band from labeling with [3H]PrBCM and [14C]-N,N'-dicyclohexylcarbodiimide (DCCD) proving its identity with the renal Na+/H+ exchanger. Our data reveal a specific interaction of PrBCM with the Na+/H+ exchanger and suggest structural relations between antiporter and receptors.

摘要

一种用于毒蕈碱型乙酰胆碱受体的共价结合标记物，丙基苯甲酰胆碱氮芥（PrBCM），可不可逆地抑制大鼠肾刷状缘膜囊泡中的Na+/H+交换体。反向转运体的底物Na+和Li+，以及抑制剂阿米洛利、5-（N-乙基-N-异丙基）阿米洛利（EIPA）和普萘洛尔，可保护反向转运体不被PrBCM灭活。用[3H]PrBCM时，一条表观分子量为65 kDa的条带被主要标记。阿米洛利可保护这条带不被[3H]PrBCM和[14C]-N，N'-二环己基碳二亚胺（DCCD）标记，证明其与肾Na+/H+交换体相同。我们的数据揭示了PrBCM与Na+/H+交换体之间的特异性相互作用，并表明反向转运体与受体之间存在结构关系。