Size of acetylcholine receptors in the membrane. An improved version of the radiation inactivation method.
The radiation inactivation method was used to study the size of acetylcholine receptors in the intact membrane-bound state. This technique was reinvestigated, and modifications were made which remove substantial difficulties affecting previous applications of it to such proteins. The molecular size was deduced here by reference to a set of protein standards: an inactivation ratio was defined relative to a given internal enzyme molecular weight standard, and a linear calibration plot for the inactivation ratios of the protein standards was constructed and applied. The acetylcholine receptor in Torpedo electric organ, cat denervated muscle, and chick embryonic muscle was found by this method to exist in the membrane as a homogeneous population of the same size in each case. This receptor, when identified thus by the alpha-neurotoxin-binding target structure, has an apparent molecular weight of 300000 or a molecular volume of about 350 nm3. In comparison, the molecular weight of the cat muscle receptor when solubilized, as analyzed by gel electrophoresis after extensive cross-linking, was found to be 270000 +/- 20000. These two values are thought to be equivalent by virtue of the situation and structure of the receptor protein in the cell membrane. If a disulfide-bridge dimeric receptor exists in the membrane (as other evidence has indicated for Torpedo), each monomer acts independently there in binding alpha-neurotoxin, since the monomers can be inactivated independently by irradiation in the Torpedo membrane. In the muscle membrane no evidence for the existence of receptor dimers, of any kind, has been found.
D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D002415:Cats; D002462:Cell Membrane / Q000378:metabolism*; D002642:Chick Embryo; D004557:Electric Organ / Q000378:metabolism; D066298:In Vitro Techniques; D008970:Molecular Weight; D009132:Muscles / Q000378:metabolism; D011487:Protein Conformation; D011950:Receptors, Cholinergic / Q000378:metabolism* / Q000528:radiation effects; D014101:Torpedo / Q000378:metabolism
D011950:Receptors, Cholinergic; D000109:Acetylcholine