Enzymatic activity is necessary for thrombin-mediated increase in endothelial permeability

Judy L. Aschner, J. M. Lennon, J. W. Fenton, Michael Aschner, A. B. Malik

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

α-Thrombin causes a dose-dependent increase in endothelial permeability as measured by the clearance rate of 125I-albumin across a monolayer of bovine pulmonary artery endothelial cells. We determined if an active catalytic site is necessary for the thrombin-mediated increase in endothelial permeability. α-Thrombin was reacted with 10-fold excess D-phenylalanyl-prolyl-arginine chloromethyl ketone (PPACK), an irreversible inhibitor that forms a covalent bond with thrombin's active site, producing an enzymatically inactive thrombin. PPACK completely inhibited the α-thrombin-mediated increase in 125I-albumin permeability. Similar results were obtained with γ-thrombin, an enzymatically active α-thrombin form with an altered fibrinogen recognition domain. PPACK alone and the active site-inhibited PPACK-α-thrombin had no effect on permeability. Diisopropylphospho (DIP)-α-thrombin was effective only in very high concentrations (10-6 M), and this effect was abolished by the addition of PPACK. These studies demonstrate that binding alone is insufficient for the thrombin-mediated increase in endothelial monolayer permeability. Thrombin's active catalytic site is a requirement for the increase in transendothelial albumin permeability.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume259
Issue number4 3-2
StatePublished - 1990
Externally publishedYes

Fingerprint

Thrombin
Permeability
Catalytic Domain
Albumins
Fibrinogen
Pulmonary Artery
Endothelial Cells
phenylalanyl-prolyl-arginine-chloromethyl ketone

Keywords

  • endothelial monolayer permeability

ASJC Scopus subject areas

  • Cell Biology
  • Physiology
  • Pulmonary and Respiratory Medicine

Cite this

Enzymatic activity is necessary for thrombin-mediated increase in endothelial permeability. / Aschner, Judy L.; Lennon, J. M.; Fenton, J. W.; Aschner, Michael; Malik, A. B.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 259, No. 4 3-2, 1990.

Research output: Contribution to journalArticle

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AU - Malik, A. B.

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