Activity and inhibition of prostasin and matriptase on apical and basolateral surfaces of human airway epithelial cells.

American Journal of Physiology. Lung Cellular and Molecular Physiology
Shilpa NimishakaviGeorge H Caughey

Abstract

Prostasin is a membrane-anchored protease expressed in airway epithelium, where it stimulates salt and water uptake by cleaving the epithelial Na(+) channel (ENaC). Prostasin is activated by another transmembrane tryptic protease, matriptase. Because ENaC-mediated dehydration contributes to cystic fibrosis (CF), prostasin and matriptase are potential therapeutic targets, but their catalytic competence on airway epithelial surfaces has been unclear. Seeking tools for exploring sites and modulation of activity, we used recombinant prostasin and matriptase to identify substrate t-butyloxycarbonyl-l-Gln-Ala-Arg-4-nitroanilide (QAR-4NA), which allowed direct assay of proteases in living cells. Comparisons of bronchial epithelial cells (CFBE41o-) with and without functioning cystic fibrosis transmembrane conductance regulator (CFTR) revealed similar levels of apical and basolateral aprotinin-inhibitable activity. Although recombinant matriptase was more active than prostasin in hydrolyzing QAR-4NA, cell surface activity resisted matriptase-selective inhibition, suggesting that prostasin dominates. Surface biotinylation revealed similar expression of matriptase and prostasin in epithelial cells expressing wild-type vs. ΔF508-mutated C...Continue Reading

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Citations

Oct 21, 2015·PloS One·Shilpa NimishakaviGeorge H Caughey
Mar 26, 2016·American Journal of Respiratory and Critical Care Medicine·James A ReihillS Lorraine Martin
Feb 27, 2017·American Journal of Physiology. Lung Cellular and Molecular Physiology·Awen MenouBruno Crestani
Apr 23, 2013·American Journal of Physiology. Lung Cellular and Molecular Physiology·Christine U Vohwinkel, István Vadász
Jun 3, 2021·International Journal of Molecular Sciences·Emma L CarrollS Lorraine Martin
Nov 3, 2021·ACS Chemical Biology·Verena Rickert-ZachariasCarsten Schultz

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