Assay of the acetylcholine receptor by affinity labeling

Arthur Karlin, Mark G. McNamee, David Cowburn

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

An accurate and sensitive assay for nicotinic acetylcholine receptor binding sites is described which is based on the specificities of receptor both for an affinity label, 4-(N-maleimido)benzyltrimethylammonium iodide (MBTA), and for α-neurotoxins from Naja venoms. It has been demonstrated that MBTA reacts exclusively with one type of subunit of the acetylcholine receptors isolated from the electric tissue of Electrophorus electricus and Torpedo californica and that this reaction is blocked in the presence of Naja naja siamensis α-neurotoxin and of other ligands of the acetylcholine binding site. Thus, in this assay the difference in the extent of labeling by MBTA in the absence and presence of N. n. siamensis toxin is considered the specific labeling of receptor. Although this assay is more complicated than direct α-neurotoxin binding, it is justified by the wellestablished site specificity of the labeling. The specific activities of several different receptor preparations determined using this assay are one-half of those determined using toxin binding. It is possible to assay accurately as little as 0.25 μg of receptor in the presence of 100-fold as much other protein.

Original languageEnglish (US)
Pages (from-to)442-451
Number of pages10
JournalAnalytical Biochemistry
Volume76
Issue number2
DOIs
StatePublished - 1976
Externally publishedYes

Fingerprint

Neurotoxins
Cholinergic Receptors
Labeling
Assays
Binding Sites
Affinity Labels
Electrophorus
Cobra Venoms
Elapidae
Torpedo
Nicotinic Receptors
Acetylcholine
Ligands
4-(N-maleimido)benzyltrimethylammonium
Tissue
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Assay of the acetylcholine receptor by affinity labeling. / Karlin, Arthur; McNamee, Mark G.; Cowburn, David.

In: Analytical Biochemistry, Vol. 76, No. 2, 1976, p. 442-451.

Research output: Contribution to journalArticle

Karlin, Arthur ; McNamee, Mark G. ; Cowburn, David. / Assay of the acetylcholine receptor by affinity labeling. In: Analytical Biochemistry. 1976 ; Vol. 76, No. 2. pp. 442-451.
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