Functional role of the cysteine 451 thiol group in the M4 helix of the gamma subunit of Torpedo californica acetylcholine receptor.
Previous chemical modification studies of the acetylcholine receptor [Yee, A.S., Corey, D.E., & McNamee, M.G. (1986) Biochemistry 25, 2110-2119] were extended by using fluorescent N-pyrenylmaleimide to alkylate purified Torpedo californica nicotinic acetylcholine receptor (AChR). Peptide sequencing of the tryptic fragments of the labeled AChR gamma subunit identified cysteines 416, 420, and 451 as the modified residues. The functional role of Cys-451 in the M4 transmembrane domain of the AChR gamma subunit was further investigated by studying the functional consequences of the site-specific mutation of this cysteine to either serine or tryptophan by using AChR mRNAs injected into Xenopus laevis oocytes. Both mutants displayed about 50% reduction in the normalized channel activity of the receptor measured as the ACh-induced conductance per femtomole of surface alpha-bungarotoxin binding sites. However, the mutations did not change other AChR functional properties such as agonist binding ability, the slow phase of desensitization, and blockade by competitive and noncompetitive antagonists. The significant reduction in AChR ion channel activity associated with the above point mutations, especially the simple change of the -SH group on Cys-451 to the -OH group, suggests that this thiol group in the M4 helix of gamma subunit may play an important role in AChR ion channel function. Previous site-directed mutations of the Cys-416 and -420 residues showed a decreased response when both of these residues were changed to phenylalanine, but not when they were changed to serine [Pradier, L., Yee, A.S., & McNamee, M.G. (1989) Biochemistry 28, 6562-6571].(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Biochemistry and Biophysics, University of California, Davis 95616.
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D000595:Amino Acid Sequence; D000818:Animals; D003545:Cysteine / Q000378:metabolism; D007473:Ion Channels / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009865:Oocytes / Q000378:metabolism; D011950:Receptors, Cholinergic / Q000235:genetics / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014101:Torpedo / Q000378:metabolism; D014982:Xenopus laevis
D007473:Ion Channels; D011950:Receptors, Cholinergic; D013438:Sulfhydryl Compounds; D003545:Cysteine