subtype differences in pre-coupling of muscarinic acetylcholine receptors毒蕈碱的乙酰胆碱受体的亚型pre-coupling差异.pdfVIP

Subtype Differences in Pre-Coupling of Muscarinic Acetylcholine Receptors ´ 1 ´ˇ ´ 1 ´ ´ 1 2 ´ ˇ 1 Jan Jakubık *, Helena Janıckova , Alena Randakova , Esam E. El-Fakahany , Vladimır Dolezal 1 Department of Neurochemistry, Institute of Physiology Academy of Sciences of the Czech Republic v.v.i., Prague, Czech Republic, 2 Department of Experimental and Clinical Pharmacology, University of Minnesota College of Pharmacy, Minneapolis, Minnesota, United States of America Abstract Based on the kinetics of interaction between a receptor and G-protein, a myriad of possibilities may result. Two extreme cases are represented by: 1/Collision coupling, where an agonist binds to the free receptor and then the agonist-receptor complex ‘‘collides’’ with the free G-protein. 2/Pre-coupling, where stable receptor/G-protein complexes exist in the absence of agonist. Pre-coupling plays an important role in the kinetics of signal transduction. Odd-numbered muscarinic acetylcholine receptors preferentially couple to Gq/11, while even-numbered receptors prefer coupling to Gi/o. We analyzed the coupling status of the various subtypes of muscarinic receptors with preferential and non-preferential G-proteins. The magnitude of receptor-G-protein coupling was determined by the proportion of receptors existing in the agonist high- affinity binding conformation. Antibodies directed against the C-terminus of the a-subunits of the individual G-proteins were used to interfere with receptor-G-protein coupling. Effects of mutations and expression level on receptor-G-protein coupling were also investigated.