Specific binding of endothelin-1 to canine tracheal epithelial cells in culture

H. Ninomiya, X. Y. Yu, Y. Uchida, S. Hasegawa, Ernst W Spannhake

Research output: Contribution to journalArticle

Abstract

We have studied the binding of endothelin-1 (ET-1) to cultured canine tracheal epithelial cells. A single specific binding site for 125I- labeled ET-1 was identified with an apparent dissociation constant (K(d)) of 0.2 nM, maximal binding sites (B(max)) of 6.7 x 103 sites/cell, and half- maximal inhibition (IC50) of 0.3 nM during a 2-h incubation period. The binding of 125I-ET-1 to these cells was inhibited by the presence of unlabeled ET-1, ET-2, or BQ-123, whereas ET-3 and sarafotoxin S6c did not compete for this binding site. These binding characteristics are consistent with those of the ETA receptor. At 37°C, specific binding continuously increased over 18 h, while at 4°C, it reached a plateau by 2 h. The increase in binding at 37°C was not associated with DNA synthesis but was dependent upon protein synthesis, suggesting that epithelial binding sites were produced continuously under these incubation conditions. Our results indicate that canine tracheal epithelial cells possess specific binding sites for ET- 1 with characteristics similar to those of the ETA receptor subtype. Because these cells are demonstrated to both release and bind ET-1, the results further suggest that ET-1 is involved in paracrine and/or autocrine control mechanisms in the airway epithelium.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume268
Issue number3 12-3
StatePublished - 1995

Fingerprint

endothelins
Endothelin-1
Canidae
cell culture
epithelial cells
Cell Culture Techniques
Epithelial Cells
binding sites
Binding Sites
dogs
receptors
cells
Inhibitory Concentration 50
inhibitory concentration 50
plateaus
epithelium
Epithelium
protein synthesis
synthesis
DNA

Keywords

  • BQ-123
  • canine airway epithelial cells
  • colchicine
  • cycloheximide
  • endothelin(A) receptor
  • primary cell culture
  • sarafotoxin S6c

ASJC Scopus subject areas

  • Cell Biology
  • Physiology
  • Pulmonary and Respiratory Medicine
  • Agricultural and Biological Sciences(all)

Cite this

Specific binding of endothelin-1 to canine tracheal epithelial cells in culture. / Ninomiya, H.; Yu, X. Y.; Uchida, Y.; Hasegawa, S.; Spannhake, Ernst W.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 268, No. 3 12-3, 1995.

Research output: Contribution to journalArticle

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AU - Spannhake, Ernst W

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