PMID: 3756139Jul 29, 1986Paper

Structural heterogeneity of the alpha subunits of the nicotinic acetylcholine receptor in relation to agonist affinity alkylation and antagonist binding

Biochemistry
M RatnamJ M Lindstrom

Abstract

The structural basis for the heterogeneity of the two agonist binding sites of the Torpedo californica acetylcholine receptor with respect to antagonist binding and reactivity toward affinity alkylating reagents was investigated. There is one agonist binding site on each of the two alpha subunits in a receptor monomer. One of these sites is easily affinity labeled with bromoacetylcholine, while more extreme conditions are required to label the other. Evidence is presented that the site which is easily labeled with bromoacetylcholine is the site with higher affinity for the antagonist d-tubocurarine. Digestion of purified alpha subunits with staphylococcal V8 protease gave two limit fragments with apparent molecular weights of 17K and 19K. Both of these fragments began at residue 46 of the alpha sequence, and both reacted with monoclonal antibodies specific for the sequence alpha 152-159 but not with antibodies specific for alpha 235-242. Their tryptic peptide maps and reactivity with a number of monoclonal antibodies were virtually identical. Only the 17-kilodalton (17-kDa) fragments stained heavily for sugars with Schiff's reagent. However, both fragments bound 125I-labeled concanavalin A. Complete removal of carbohydrate dete...Continue Reading

Citations

Jan 1, 1987·Molecular Neurobiology·J LindstromP Whiting
Sep 11, 1992·Brain Research. Molecular Brain Research·R H LoringK J Stevenson
Dec 24, 1997·Brain Research. Brain Research Reviews·H R Arias
Feb 28, 2001·Proceedings of the National Academy of Sciences of the United States of America·P T WilliamsonA Watts
May 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·B ChatrenetA Ménez
Mar 1, 1991·The Journal of Cell Biology·M S SaediJ Lindstrom
Aug 1, 1986·Psychological Medicine·L BrosanM Broadbent

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