Purification and characterization of nicotinic acetylcholine receptors from muscle

George Kemp, Barbara Morley, Donard Dwyer, Ronald J. Bradley

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The nicotinic acetylcholine receptor was purified from normal and denervated rat skeletal muscle. The purification protocol included αcobratoxin biospecific adsorption, ion exchange chromatography, and gel filtration steps. The highest specific activity achieved was 7.5 pmol of 125I-αbungarotoxin binding sites per μg protein. Sodium dodecyl sulfate gel electrophoresis of purified AChR revealed subunits with molecular weights of 42,000 and 66,000 daltons and a minor component with a molecular weight of 52,000 daltons. Normal muscle AChR is comprised of one toxin binding component. Upon denervation a second component appears, but both components are increased as a consequence of denervation. A dissociation constant of 1.5 × 10-8M was determined for dtubocurarine from receptor from both normal and denervated muscle. A dissociation constant of 1 × 10-7M for acetylcholine, perhaps analogous to the high affinity acetylcholine binding observed in electric fish receptor, was determined.

Original languageEnglish (US)
Pages (from-to)229-257
Number of pages29
JournalMolecular Membrane Biology
Volume3
Issue number3
DOIs
StatePublished - Jan 1 1980

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Nicotinic Receptors
Denervation
Acetylcholine
Purification
Muscle
Molecular Weight
Electric Fish
Bungarotoxins
Muscles
Ion Exchange Chromatography
Gels
Molecular weight
Sodium Dodecyl Sulfate
Adsorption
Gel Chromatography
Electrophoresis
Skeletal Muscle
Binding Sites
Chromatography
Fish

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Purification and characterization of nicotinic acetylcholine receptors from muscle. / Kemp, George; Morley, Barbara; Dwyer, Donard; Bradley, Ronald J.

In: Molecular Membrane Biology, Vol. 3, No. 3, 01.01.1980, p. 229-257.

Research output: Contribution to journalArticle

Kemp, George ; Morley, Barbara ; Dwyer, Donard ; Bradley, Ronald J. / Purification and characterization of nicotinic acetylcholine receptors from muscle. In: Molecular Membrane Biology. 1980 ; Vol. 3, No. 3. pp. 229-257.
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