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 language | English (US) |
|---|---|
| Pages (from-to) | 229-257 |
| Number of pages | 29 |
| Journal | Molecular Membrane Biology |
| Volume | 3 |
| Issue number | 3 |
| DOIs | |
| State | Published - Jan 1 1980 |
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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 journal › Article
}
TY - JOUR
T1 - Purification and characterization of nicotinic acetylcholine receptors from muscle
AU - Kemp, George
AU - Morley, Barbara
AU - Dwyer, Donard
AU - Bradley, Ronald J.
PY - 1980/1/1
Y1 - 1980/1/1
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=0019303153&partnerID=8YFLogxK
U2 - 10.3109/09687688009063887
DO - 10.3109/09687688009063887
M3 - Article
C2 - 7382840
AN - SCOPUS:0019303153
VL - 3
SP - 229
EP - 257
JO - Molecular Membrane Biology
JF - Molecular Membrane Biology
SN - 0968-7688
IS - 3
ER -