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Molecular Pharmacology

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Abstract

Internalization of endothelin by cultured human vascular smooth muscle cells: characterization and physiological significance.

T J Resink, T Scott-Burden, C Boulanger, E Weber and F R Bühler
Molecular Pharmacology August 1990, 38 (2) 244-252;
T J Resink
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T Scott-Burden
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C Boulanger
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E Weber
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F R Bühler
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Abstract

The binding and internalization of 125I-endothelin (125I-ET-1) was studied in cultured human vascular smooth muscle cells (hVSMC). Discrimination between surface-bound and internalized radiolabeled ligand was achieved using either acetic acid or trypsin treatment of cell layers, with the two procedures yielding comparable results. Total cellular 125I-ET-1 binding hVSMC at 37 degrees was rapid and reached near equilibrium within 30 min. Such binding could be resolved into surface-bound (acid/trypsin-sensitive) and internalized (acid/trypsin-resistant) components. The accumulation of internalized 125I-ET-1 was temperature dependent and occurred at 37 degrees (t1/2 approximately 15 min) but not at 4 degrees. Internalization of 125I-ET-1 by hVSMC was reversibly inhibited by the transglutaminase inhibitor dansylcadaverine (half-maximal inhibitory concentration, approximately 400 microM). Cytosolic acidification of hVSMC (from pH approximately 6.8 to approximately 6.3) by incubation with potassium acetate in a choline buffer also inhibited 125I-ET-1 internalization. Our observation indicate that smooth muscle cells internalize ET-1 via the clathrin-mediated endocytotic pathway. Dansylcadaverine and other inhibitors of transglutaminase inhibited ET-1-stimulated inositol phospholipid hydrolysis in hVSMC and decreased ET-1-induced vasoconstriction in isolated endothelium-denuded blood vessels. Internalization of ET-1 may, therefore, be relevant to the characteristically protracted physiological effects of this peptide on the vasculature.

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Molecular Pharmacology
Vol. 38, Issue 2
1 Aug 1990
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Abstract

Internalization of endothelin by cultured human vascular smooth muscle cells: characterization and physiological significance.

T J Resink, T Scott-Burden, C Boulanger, E Weber and F R Bühler
Molecular Pharmacology August 1, 1990, 38 (2) 244-252;

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Abstract

Internalization of endothelin by cultured human vascular smooth muscle cells: characterization and physiological significance.

T J Resink, T Scott-Burden, C Boulanger, E Weber and F R Bühler
Molecular Pharmacology August 1, 1990, 38 (2) 244-252;
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