Direct photoaffinity labeling of the high affinity nitrendipine-binding site in subcellular membrane fractions isolated from canine myocardium

Kevin P Campbell; Greg M Lipshutz; Gerald H Denney

doi:10.1016/S0021-9258(18)91018-5

$Direct photoaffinity labeling of the high affinity nitrendipine-binding site in subcellular membrane fractions isolated from canine myocardium$

Journal article

Open access

Peer reviewed

Direct photoaffinity labeling of the high affinity nitrendipine-binding site in subcellular membrane fractions isolated from canine myocardium

Kevin P Campbell, Greg M Lipshutz and Gerald H Denney

The Journal of biological chemistry, Vol.259(9), pp.5384-5387

1984

DOI: 10.1016/S0021-9258(18)91018-5

PMID: 6325434

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Abstract

[3H]Nitrendipine and high intensity ultraviolet irradiation have been used to photoaffinity label the protein component of the high affinity nitrendipine-binding site in subcellular membrane fractions from canine cardiac muscle. Irradiation of isolated cardiac membranes in the presence of [3H]nitrendipine resulted in the covalent labeling of a protein component that migrated on sodium dodecyl sulfate-polyacrylamide gels with an apparent molecular weight of 32,000. Incorporation of [3H]nitrendipine did not occur in the absence of irradiation. The photoaffinity labeling of the 32,000-Da protein by [3H]nitrendipine was inhibited by excess unlabeled nitrendipine, nifedipine, or verapamil. EDTA, ATP, and La3+, which are known to reduce high affinity nitrendipine binding, also inhibited the photoaffinity labeling of this membrane protein by [3H]nitrendipine. The 32,000-Da [3H]nitrendipine-labeled protein was found to be enriched in the ryanodine-sensitive fraction of cardiac sarcoplasmic reticulum and absent from the ryanodine-insensitive fraction of cardiac sarcoplasmic reticulum which is known to lack high affinity nitrendipine binding. Therefore, the 32,000-Da photoaffinity-labeled [3H]nitrendipine-binding protein exhibits properties identical to those expected for the protein component of the high affinity nitrendipine-binding site in isolated cardiac membranes.

Fundamental and applied biological sciences. Psychology

Biological and medical sciences

Molecular and cellular biology

Cell structures and functions

Cell membranes. Ionic channels. Membrane pores

Details

Title: Subtitle: Direct photoaffinity labeling of the high affinity nitrendipine-binding site in subcellular membrane fractions isolated from canine myocardium
Creators: Kevin P Campbell - Univ. Iowa, dep. physiologu biophysics, Iowa City IA 52242, United States
Greg M Lipshutz - Univ. Iowa, dep. physiologu biophysics, Iowa City IA 52242, United States
Gerald H Denney - Univ. Iowa, dep. physiologu biophysics, Iowa City IA 52242, United States
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.259(9), pp.5384-5387
DOI: 10.1016/S0021-9258(18)91018-5
PMID: 6325434
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: American Society for Biochemistry and Molecular Biology; Bethesda, MD
Language: English
Date published: 1984
Academic Unit: Neurology; Molecular Physiology and Biophysics; Iowa Neuroscience Institute
Record Identifier: 9984068262502771

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