Endothelin-receptor interactions. Role of a putative sulfhydryl on the endothelin receptor

Michael J. Spinella; Rebecca Kottke; Harold I. Magazine; Matthew S. Healy; John A. Catena; Philip Wilken; Thomas T. Andersen

doi:10.1016/0014-5793(93)80970-6

Endothelin-receptor interactions. Role of a putative sulfhydryl on the endothelin receptor

Michael J. Spinella, Rebecca Kottke, Harold I. Magazine, Matthew S. Healy, John A. Catena, Philip Wilken, Thomas T. Andersen

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

Abstract

The mechanism of action of endothelin-receptor interactions was studied, using radioligand binding assays and SDS-PAGE, to investigate the possibility of disulfide interchange. Electrophoretic analysis suggested involvement of disulfide bond(s) in the receptor-ligand complex. Treatment of Et receptors with sulfhydryl-specific alkylating reagents (NEM or others) resulted in decreased ability to bind [¹²⁵I]Et-1. [Dpr¹-Asp¹⁵]Et-1, an antagonist homologous to Et but with an amide link replacing one of the disulfides, bound to Et receptors reversibly, but binding of Et-1 was less reversible. Preincubation of receptors with Et-L, but not with [Dpr¹-Asp¹⁵]Et-L, protected receptors from alkylation with [¹⁴C]NEM. The data suggest that the Et receptor has a sulfhydryl group at or near the Et binding site. A model is proposed in which the role of the putative sulfhydryl group is discussed.

Original language	English (US)
Pages (from-to)	82-88
Number of pages	7
Journal	FEBS Letters
Volume	328
Issue number	1-2
DOIs	https://doi.org/10.1016/0014-5793(93)80970-6
State	Published - Aug 9 1993
Externally published	Yes

Keywords

Antagonist
Eendothelin receptor
Endothelin
Mechanism

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

Online availability

10.1016/0014-5793(93)80970-6

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title = "Endothelin-receptor interactions. Role of a putative sulfhydryl on the endothelin receptor",

abstract = "The mechanism of action of endothelin-receptor interactions was studied, using radioligand binding assays and SDS-PAGE, to investigate the possibility of disulfide interchange. Electrophoretic analysis suggested involvement of disulfide bond(s) in the receptor-ligand complex. Treatment of Et receptors with sulfhydryl-specific alkylating reagents (NEM or others) resulted in decreased ability to bind [125I]Et-1. [Dpr1-Asp15]Et-1, an antagonist homologous to Et but with an amide link replacing one of the disulfides, bound to Et receptors reversibly, but binding of Et-1 was less reversible. Preincubation of receptors with Et-L, but not with [Dpr1-Asp15]Et-L, protected receptors from alkylation with [14C]NEM. The data suggest that the Et receptor has a sulfhydryl group at or near the Et binding site. A model is proposed in which the role of the putative sulfhydryl group is discussed.",

keywords = "Antagonist, Eendothelin receptor, Endothelin, Mechanism",

author = "Spinella, {Michael J.} and Rebecca Kottke and Magazine, {Harold I.} and Healy, {Matthew S.} and Catena, {John A.} and Philip Wilken and Andersen, {Thomas T.}",

note = "Funding Information: Ackno,l,lellgernents This research was supported by the Margaret C. Finnerty Ahard in Cardtovascular Research (Albany Medical College), American Heart Association New York State Affiliate award AHA 89-0256, and American Heart Associatron (National) award 920 11500.",

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AU - Spinella, Michael J.

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AU - Healy, Matthew S.

AU - Catena, John A.

AU - Wilken, Philip

AU - Andersen, Thomas T.

N1 - Funding Information: Ackno,l,lellgernents This research was supported by the Margaret C. Finnerty Ahard in Cardtovascular Research (Albany Medical College), American Heart Association New York State Affiliate award AHA 89-0256, and American Heart Associatron (National) award 920 11500.

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N2 - The mechanism of action of endothelin-receptor interactions was studied, using radioligand binding assays and SDS-PAGE, to investigate the possibility of disulfide interchange. Electrophoretic analysis suggested involvement of disulfide bond(s) in the receptor-ligand complex. Treatment of Et receptors with sulfhydryl-specific alkylating reagents (NEM or others) resulted in decreased ability to bind [125I]Et-1. [Dpr1-Asp15]Et-1, an antagonist homologous to Et but with an amide link replacing one of the disulfides, bound to Et receptors reversibly, but binding of Et-1 was less reversible. Preincubation of receptors with Et-L, but not with [Dpr1-Asp15]Et-L, protected receptors from alkylation with [14C]NEM. The data suggest that the Et receptor has a sulfhydryl group at or near the Et binding site. A model is proposed in which the role of the putative sulfhydryl group is discussed.

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Endothelin-receptor interactions. Role of a putative sulfhydryl on the endothelin receptor

Abstract

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