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PMID: 6329680 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The rotational diffusion of the acetylcholine receptor in Torpeda marmorata membrane fragments studied with a spin-labelled alpha-toxin: importance of the 43 000 protein(s).

The EMBO journal ·Vol. 1 ·No. 4 ·1982-00-00 ·Pages 439-45

Rousselet A, Cartaud J, Devaux PF, Changeux JP

Abstract

The rotational diffusion of the acetylcholine (ACh) receptor in subsynaptic membrane fragments from Torpedo marmorata electric organ was investigated with a spin-labelled alpha-bungarotoxin. A toxin with two spin labels was first synthesized; the conventional electron spin resonance spectrum (e.s.r.) of this toxin bound to the receptor indicated: (1) a complete immobilization of the probes; and (2) a strong spin-spin interaction that was not, or barely, seen in solution. The modification of the degree of spin-spin interaction is taken as an indication of a toxin conformational change accompanying its binding to the ACh-receptor. To avoid spin-spin interaction a single-labelled toxin was made and used to follow the rotational diffusion of the receptor by saturation transfer e.s.r. (ST-e.s.r.). With native membranes a high immobilization of the ACh-receptor was noticed. Reduction of the membranes by dithiothreitol had little effect on this motion. Only extraction of the 43 000 protein(s) by pH 11 treatment was able to enhance the rotational diffusion of the ACh-receptor protein (rotational correlation time by ST-e.s.r. in the 0.5 - 1 X 10(-4) s range) and to allow its lateral diffusion in the plane of the membrane fragments (observed by electron microscopy after freeze-etching or negative staining).

MeSH Terms
Animals Bungarotoxins/metabolism Cell Membrane/physiology,ultrastructure Diffusion Electric Organ/physiology Electron Spin Resonance Spectroscopy Microscopy, Electron Molecular Weight Receptors, Cholinergic/isolation & purification,physiology Torpedo
Chemicals
Bungarotoxins Receptors, Cholinergic
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Rousselet A
Cartaud J
Devaux P F
Changeux J P
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39 references, click to expand
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1982-00-00
Pages
439-45
Language
English
Region
England
NLM ID
8208664
PMCID
PMC553065
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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