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In vitro effects of reactive O2 species on the β-receptor-adenylyl cyclase system

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Abstract

The irreversible loss of activity of the sarcolemma-localized β-receptor-adenylyl cyclase system (β-RAS) in myocardial ischemia is a well documented phenomenon. Alterations in the sarcolemma (SL) induced by reactive O2 species could be responsible for this loss. Therefore the influence of oxidation of SH-groups and lipid peroxidation induced by Fe2+/Vit. C on the β-RAS activity was studied. During incubation of SL with Fe2+/Vit. C a transient enhancement followed by a continuous loss of the β-RAS activity (isoprenaline-, NaF-, Gpp(NH)p-, forskolin-stimulated and basal activity) was observed. In contrast there occurred a continuous loss of SH-groups and lipid peroxidation, beginning immediately after the start of incubation. Loss of SH-groups and lipid peroxidation as well as changes in the β-RAS did not take place in the presence of the antioxidant t-Butyl-4-hydroxyanisole (BHA) or the Fe2+-chelator EGTA. In view of the known ischemia-induced formation of reactive O2 species our results show that these powerful oxidants could contribute to the modulation of the β-RAS during myocardial ischemia.

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Abbreviations

BHA:

t-Butyl-4-Hydroxyanisole

DTNB:

5,5′-Dithio-bis-(2-nitrobenzoic acid)

DTT:

Dithiothreitol

EGTA:

Ethyleneglycol-bis-(2-amino-ethylether)-N,N,N,N′-Tetraacetic Acid

Gpp(NH)p:

Guanylylimidodiphosphate

HSA:

Human Serum Albumine

PMSF:

Phenylmethylsulfonyl Fluoride

β-RAS:

β-Receptor-Adenylyl cyclase System

SDS:

Sodium Dodecylsulfate

TRIS:

Tris(hydroxymethyl)-aminomethane

SL:

Sarcolemma

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Authors and Affiliations

  1. Institut für Herz-Kreislauf-Forschung, Berlin, Germany

    Liane Will-Shahab & Irmgard Küttner

  2. Institut für Pathologische und Klinische Biochemie, Humboldt-Universität zu Berlin, Schumannstr. 20/21, D-1040, Berlin, Germany

    Ingolf Schimke, Annekathrin Haberland, Liane Will-Shahab, Irmgard Küttner & Bärbel Papies

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  1. Ingolf Schimke
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  2. Annekathrin Haberland
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  3. Liane Will-Shahab
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  4. Irmgard Küttner
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  5. Bärbel Papies
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Schimke, I., Haberland, A., Will-Shahab, L. et al. In vitro effects of reactive O2 species on the β-receptor-adenylyl cyclase system. Mol Cell Biochem 110, 41–46 (1992). https://doi.org/10.1007/BF02385004

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  • DOI: https://doi.org/10.1007/BF02385004

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