Subcellular Distribution and Isolation of the Ca2+ Antagonist Receptor Associated with the Voltage Regulated Ca2+ Channel from Rabbit Heart Muscle

Overview

Journal Mol Cell Biochem

Publisher Springer

Specialty Biochemistry

Date 1987 Aug 1

PMID 2444872

Citations 4

Authors

B S Tuana

B J Murphy

Q Yi

Affiliations

Soon will be listed here.

Abstract

The Ca2+ antagonist binding sites associated with the voltage dependent calcium channel in rabbit myocardium were found to distribute with the sarcolemmal Na+ + K+ ATPase and adenylate cyclase activities during subcellular fractionation on sucrose-density gradients. The equilibrium dissociation constants (KD) for the binding of [3H]nitrendipine and [3H]verapamil were 0.31 +/- 0.04 nM and 4.1 +/- 0.5 nM respectively, and displayed an average density of 0.55 +/- 0.05 pmol/mg and 0.4 +/- 0.03 pmol/mg protein respectively for the most enriched membrane fraction. The Ca2+ antagonist binding sites were solubilized from the membranes with the detergent 3-[(3-cholamidopropyl)dimethylammonio]propanesulfonate, and specific binding sites for [3H]PN200-110, [3H]verapamil and [3H]diltiazem were isolated on a wheat-germ lectin column. The binding sites for [3H]PN200-110 were enriched about 2,500 fold as compared with the original homogenate and displayed a density of 28.5 +/- 8 pmole/mg protein in the isolated fraction. Sodium dodecyl sulfate gel electrophoresis of the isolated drug binding proteins indicated enrichment of proteins of Mr 170,000, 140,000, 130,000, 100,000 and 53,000. The isolated receptor contained an intrinsic kinase activity that phosphorylated glycoproteins of Mr 170,000 and 53,000. Exogenously added cAMP-kinase stimulated phosphorylation of the 170,000, 100,000, 53,000 and 28,000 Mr glycoproteins in the receptor fraction. The results of this study indicate that the binding sites for [3H]nitrendipine, [3H]PN200-110, [3H]verapamil and [3H]diltiazem residue on glycoprotein(s) which are of sarcolemmal origin, and co-purify together on wheat germ lectin columns. The polypeptide composition of the Ca2+ antagonist binding sites from cardiac muscle appears to be very similar to that of the dihydropyridine receptor in skeletal muscle.

Citing Articles

Ca2+-antagonists inhibit the N-methyltransferase-dependent synthesis of phosphatidylcholine in the heart.

Tappia P, Okumura K, Kawabata K, Shah K, Nijjar M, Panagia V Mol Cell Biochem. 2001; 221(1-2):89-98.

PMID: 11506191 DOI: 10.1023/a:1010905221770.

Dystrophin predominantly localizes to the transverse tubule/Z-line regions of single ventricular myocytes and exhibits distinct associations with the membrane.

Peri V, Ajdukovic B, Holland P, Tuana B Mol Cell Biochem. 1994; 130(1):57-65.

PMID: 8190121 DOI: 10.1007/BF01084268.

A calmodulin dependent protein kinase activity associated with rabbit heart sarcolemma.

Tuana B, Murphy B, Schwarzkopf C Mol Cell Biochem. 1987; 78(1):47-54.

PMID: 3454868 DOI: 10.1007/BF00224423.

The purified Ca2+ antagonist receptor from skeletal muscle: subunit structure, photoaffinity labeling and endogenous protein kinase activity.

Tuana B, Murphy B, Yi Q Mol Cell Biochem. 1988; 80(1-2):133-43.

PMID: 2845255 DOI: 10.1007/BF00231011.

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