Monoclonal antibodies that coimmunoprecipitate the 1,4-dihydropyridine and phenylalkylamine receptors and reveal the Ca/sup 2 +/ channel structure
Monoclonal hybridoma cell lines secreting antibodies against the (+)-PN 200-110 and the (-)-demethoxyverapamil binding components of the voltage-dependent calcium channel from rabbit transverse-tubule membranes have been isolated. The specificity of these monoclonal antibodies was established by their ability to coimmunoprecipitate (+)-(/sup 3/H)PN 200-110 and (-)-(/sup 3/H)demethoxyverapamil receptors. Monoclonal antibodies described in this work cross-reacted with rat, mouse, chicken, and frog skeletal muscle Ca/sup 2 +/ channels but not with crayfish muscle Ca/sup 2 +/ channels. Cross-reactivity was also detected with membranes prepared from rabbit heart, brain, and intestinal smooth muscle. These antibodies were used in immunoprecipitation experiments with /sup 125/I-labeled detergent (3-((3-cholamidopropyl)dimethylammonio)-1-propanesulfonate (CHAPS) and digitonin) solubilized membranes. They revealed a single immunoprecipitating component of molecular weight (M/sub r/) 170,000 in nonreducing conditions. After disulfide bridge reduction the CHAPS-solubilized (+)-PN 200-110-(-)-demethoxyverapamil binding component gave rise to a large peptide of M/sub r/ 140,000 and to smaller polypeptides of M/sub r/ 30,000 and 26,000 whereas the digitonin-solubilized receptor appeared with subunits at M/sub r/ 170,000, 140,000, 30,000, and 26,000. All these results taken together are interpreted as showing that both the 1,4-dihydropyridine and the phenylalkylamine receptors are part of a single polypeptide chain of M/sub r/ 170,000.
- Research Organization:
- Centre de Biochimie du CNRS, Nice, France
- OSTI ID:
- 6524122
- Journal Information:
- Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 26:1; ISSN BICHA
- Country of Publication:
- United States
- Language:
- English
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62 RADIOLOGY AND NUCLEAR MEDICINE
ADDITIVES
ALKALINE EARTH METAL COMPOUNDS
ANIMALS
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