Affinity alkylation labels two subunits of the reduced acetylcholine receptor from mammalian muscle.

Abstract
The acetylcholine receptor from denervated rat skeletal muscle was purified by affinity chromatography and, after reduction, was treated with the affinity alkylating agent 4-(N-maleimido)benzyltri[3H]methylammonium iodide. The receptor specifically incorporated approximately 1 mol of alkylating agent/mol of 125I-labeled .alpha.-bungarotoxin bound. Analysis of the labeled receptor by polyacrylamide gel electrophoresis in sodium dodecyl sulfate showed that 2 subunits were labeled; their apparent MW were 45,000 and 49,000. The affinity reagent labeled a 2nd site for acetylcholine binding in the muscle receptor that was not labeled in receptors from Electrophorus or Torpedo.

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