High affinity specific [3H] (+)PN 200-110 binding to dihydropyridine receptors associated with calcium channels in rat cerebral cortex and heart

Howard R. Lee; William R. Roeske; Henry I. Yamamura

doi:10.1016/0024-3205(84)90340-0

High affinity specific [³H] (+)PN 200-110 binding to dihydropyridine receptors associated with calcium channels in rat cerebral cortex and heart

Howard R. Lee, William R. Roeske, Henry I. Yamamura

Medicine

Research output: Contribution to journal › Article › peer-review

102 Scopus citations

Abstract

The binding properties of the 1,4-dihydropyridine calcium channel antagonist, [³H](⁺)PN 200-110, were studied in rat cerebral cortical and cardiac homogenates (37°C, Krebs phosphate buffer). Specific binding of [³H](⁺)PN 200-110 was saturable, reversible, and of high affinity (K_d values are 35 and 64 pM for the cerebral cortex and heart, respectively). In parallel studies with [³H](⁺)PN 200-110, the dissociation constant of [³H]nitrendipine was 10-12 times higher. Substituted dihydropyridine calcium channel antagonists and agonists competitively inhibited specific [³H](⁺)PN 200-110 binding, but d-cis diltiazem enhanced and verapamil incompletely inhibited [³H](⁺)PN 200-110 binding in both the cerebral cortex and the heart. The effects of diltiazem and verapamil on [³H](⁺)PN 200-110 binding were due mainly to alterations in the dissociation constant (K_d), without alterations in the binding density (B_max). The new [³H](⁺)PN 200-110 receptor binding assay is remarkable for its low degree of nonspecific binding as compared to [³H]nitrendipine at physiological temperatures. [³H(⁺)PN 200-110 is a useful ligand for the further analysis of the dihydropyridine binding sites associated with calcium channels.

Original language	English (US)
Pages (from-to)	721-732
Number of pages	12
Journal	Life Sciences
Volume	35
Issue number	7
DOIs	https://doi.org/10.1016/0024-3205(84)90340-0
State	Published - Aug 13 1984

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Pharmacology, Toxicology and Pharmaceutics(all)

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@article{a3487ffada6743cb886de1a07eb8e890,

title = "High affinity specific [3H] (+)PN 200-110 binding to dihydropyridine receptors associated with calcium channels in rat cerebral cortex and heart",

abstract = "The binding properties of the 1,4-dihydropyridine calcium channel antagonist, [3H](+)PN 200-110, were studied in rat cerebral cortical and cardiac homogenates (37°C, Krebs phosphate buffer). Specific binding of [3H](+)PN 200-110 was saturable, reversible, and of high affinity (Kd values are 35 and 64 pM for the cerebral cortex and heart, respectively). In parallel studies with [3H](+)PN 200-110, the dissociation constant of [3H]nitrendipine was 10-12 times higher. Substituted dihydropyridine calcium channel antagonists and agonists competitively inhibited specific [3H](+)PN 200-110 binding, but d-cis diltiazem enhanced and verapamil incompletely inhibited [3H](+)PN 200-110 binding in both the cerebral cortex and the heart. The effects of diltiazem and verapamil on [3H](+)PN 200-110 binding were due mainly to alterations in the dissociation constant (Kd), without alterations in the binding density (Bmax). The new [3H](+)PN 200-110 receptor binding assay is remarkable for its low degree of nonspecific binding as compared to [3H]nitrendipine at physiological temperatures. [3H(+)PN 200-110 is a useful ligand for the further analysis of the dihydropyridine binding sites associated with calcium channels.",

author = "Lee, {Howard R.} and Roeske, {William R.} and Yamamura, {Henry I.}",

note = "Funding Information: This study was supported by a grant from the American Heart Asociation, Arizona Affiliate, and by USPHS grants HL-29565, MH-27257, and MH-30626. H.R.L. is supported by a clinical pharmacology fellowship from the Pharmaceutical Manufacturers Association Foundation. W.R.R. is a recipient of a USPHS Research Career Development Award (HL-00776) from the National Heart Lung and Blood Institute. H.I.Y. is a recipient of a USPHS Research Scientist Development Award, Type II (MH-O0095) from the National Insititute of Mental Health. We thank Tom Toneff and Martha Ackerman for their technical assistance and Pat Gonzalez for her secretarial assistance.",

year = "1984",

month = aug,

day = "13",

doi = "10.1016/0024-3205(84)90340-0",

language = "English (US)",

volume = "35",

pages = "721--732",

journal = "Life Sciences",

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number = "7",

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T1 - High affinity specific [3H] (+)PN 200-110 binding to dihydropyridine receptors associated with calcium channels in rat cerebral cortex and heart

AU - Lee, Howard R.

AU - Roeske, William R.

AU - Yamamura, Henry I.

N1 - Funding Information: This study was supported by a grant from the American Heart Asociation, Arizona Affiliate, and by USPHS grants HL-29565, MH-27257, and MH-30626. H.R.L. is supported by a clinical pharmacology fellowship from the Pharmaceutical Manufacturers Association Foundation. W.R.R. is a recipient of a USPHS Research Career Development Award (HL-00776) from the National Heart Lung and Blood Institute. H.I.Y. is a recipient of a USPHS Research Scientist Development Award, Type II (MH-O0095) from the National Insititute of Mental Health. We thank Tom Toneff and Martha Ackerman for their technical assistance and Pat Gonzalez for her secretarial assistance.

PY - 1984/8/13

Y1 - 1984/8/13

N2 - The binding properties of the 1,4-dihydropyridine calcium channel antagonist, [3H](+)PN 200-110, were studied in rat cerebral cortical and cardiac homogenates (37°C, Krebs phosphate buffer). Specific binding of [3H](+)PN 200-110 was saturable, reversible, and of high affinity (Kd values are 35 and 64 pM for the cerebral cortex and heart, respectively). In parallel studies with [3H](+)PN 200-110, the dissociation constant of [3H]nitrendipine was 10-12 times higher. Substituted dihydropyridine calcium channel antagonists and agonists competitively inhibited specific [3H](+)PN 200-110 binding, but d-cis diltiazem enhanced and verapamil incompletely inhibited [3H](+)PN 200-110 binding in both the cerebral cortex and the heart. The effects of diltiazem and verapamil on [3H](+)PN 200-110 binding were due mainly to alterations in the dissociation constant (Kd), without alterations in the binding density (Bmax). The new [3H](+)PN 200-110 receptor binding assay is remarkable for its low degree of nonspecific binding as compared to [3H]nitrendipine at physiological temperatures. [3H(+)PN 200-110 is a useful ligand for the further analysis of the dihydropyridine binding sites associated with calcium channels.

AB - The binding properties of the 1,4-dihydropyridine calcium channel antagonist, [3H](+)PN 200-110, were studied in rat cerebral cortical and cardiac homogenates (37°C, Krebs phosphate buffer). Specific binding of [3H](+)PN 200-110 was saturable, reversible, and of high affinity (Kd values are 35 and 64 pM for the cerebral cortex and heart, respectively). In parallel studies with [3H](+)PN 200-110, the dissociation constant of [3H]nitrendipine was 10-12 times higher. Substituted dihydropyridine calcium channel antagonists and agonists competitively inhibited specific [3H](+)PN 200-110 binding, but d-cis diltiazem enhanced and verapamil incompletely inhibited [3H](+)PN 200-110 binding in both the cerebral cortex and the heart. The effects of diltiazem and verapamil on [3H](+)PN 200-110 binding were due mainly to alterations in the dissociation constant (Kd), without alterations in the binding density (Bmax). The new [3H](+)PN 200-110 receptor binding assay is remarkable for its low degree of nonspecific binding as compared to [3H]nitrendipine at physiological temperatures. [3H(+)PN 200-110 is a useful ligand for the further analysis of the dihydropyridine binding sites associated with calcium channels.

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U2 - 10.1016/0024-3205(84)90340-0

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JO - Life Sciences

JF - Life Sciences

SN - 0024-3205

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