What question did this study set out to answer?

To solubilize, partially purify, and characterize the physical and ligand-binding properties of nicotinic acetylcholine receptors from rat cerebral cortices.

October 1, 1976Journal of Biological ChemistryOpen Access

Nicotinic acetylcholine receptor from rat brain. Solubilization, partial purification, and characterization.

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Authors

PSPaul M. SalvaterraCity Of Hope National Medical Center HMH.R. MahlerNational Institutes of Health

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Implication

Biochemical study demonstrates successful isolation and characterization of nicotinic acetylcholine receptors in rat brain tissue, indicating distinct molecular mass and ligand affinity.

Key Points

To solubilize, partially purify, and characterize the physical and ligand-binding properties of nicotinic acetylcholine receptors from rat cerebral cortices.
Extracted crude membrane fractions from rat cerebral cortices using the nonionic detergent Triton X-100.
Partially purified the receptor using Naja naja siamensis alpha-toxin coupled to Sepharose 4B affinity chromatography.
Assessed binding kinetics with 125I-labeled alpha-bungarotoxin and determined hydrodynamic parameters using sucrose gradient sedimentation and gel filtration.
Binding of alpha-bungarotoxin exhibited a forward rate constant of 0.38 x 10^5 M^-1 s^-1 and a dissociation rate constant of 1.23 x 10^-5 s^-1 (half-life of 15.6 hours), yielding a dissociation constant (Kd) of 3.24 x 10^-10 M.
Inhibition assays confirmed that various cholinergic ligands selectively blocked toxin binding, verifying nicotinic specificity.
Hydrodynamic evaluation established a sedimentation coefficient of 12.9 S, a Stokes radius of 7.5 nm, and a detergent-corrected molecular weight of 357,000.

Cite This Study

Salvaterra et al. (1976) studied this question.

synapsesocial.com/papers/6a71fc272163a0a01bc5e12f https://doi.org/10.1016/s0021-9258(20)81863-8

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Implication

Biochemical study demonstrates successful isolation and characterization of nicotinic acetylcholine receptors in rat brain tissue, indicating distinct molecular mass and ligand affinity.

Nicotinic acetylcholine receptor from rat brain. Solubilization, partial purification, and characterization.

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Key Points

Cite This Study

Nicotinic acetylcholine receptor from rat brain. Solubilization, partial purification, and characterization.

Discussion

Member takes

Implication

Key Points

Cite This Study

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