Protease effects on the structure of acetylcholine receptor membranes from Torpedo californica.
Open Access
- 1 June 1980
- journal article
- research article
- Published by Rockefeller University Press in The Journal of cell biology
- Vol. 85 (3), 823-838
- https://doi.org/10.1083/jcb.85.3.823
Abstract
Protease digestion of acetylcholine receptor-rich membranes derived from T. californica electroplaques by homogenization and isopycnic centrifugation results in degradation of all receptor subunits without any significant effect on the appearance in electron micrographs, the toxin binding ability or the sedimentation value of the receptor molecule. Such treatment does produce dramatic changes in the morphology of the normally 0.5-2 .mu.m diameter spherical vesicles when observed by either negative-stain or freeze-fracture EM. Removal of peripheral, apparently nonreceptor polypeptides by alkali stripping results in increased sensitivity of the acetylcholine receptor membranes to the protease trypsin as indicated by SDS gel electrophoretic patterns and by the extent of morphologic change observed in vesicle structure. Trypsin digestion of alkali-stripped receptor membranes results in a limit degradation pattern of all 4 receptor subunits, whereupon all the vesicles undergo the morphological transformation to minivesicles. The protein-induced morphological transformation and the limit digestion pattern of receptor membranes are unaffected by whether the membranes are prepared so as to preserve the receptor as a disulfide bridged dimer, or prepared so as to generate monomeric receptor.This publication has 37 references indexed in Scilit:
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