Protease inhibitors suppress the formation of tight junctions in gastrointestinal cell lines.
Tight junctions (TJ) of the fascia occludens type can be induced in the human colon adenocarcinoma cell lines HT29 and Caco-2 by treatment with 320 mM cesium sulfate. This process can be completely inhibited by the protease inhibitors leupeptin and antipain. The concentration for 50% inhibition was 32 microM leupeptin and 270 microM antipain, respectively. In the polarized colon carcinoma cell line Caco-2, the spontaneous formation of histotypical TJ and the development of transepithelial electrical resistance do not occur when the cells are cultured in medium containing 400 microM leupeptin. Following the removal of leupeptin, zonula occludens type TJ and electrical resistance develop synchronously during a period of 4 h. Dihydroleupeptin, the alcohol analog of leupeptin, inhibits neither the spontaneous nor the induced assembly of TJ fibrils. Thus, the aldehyde group of leupeptin is essential for activity. These data suggest that the salt-induced as well as the spontaneous formation of TJ involve cellular proteases which are susceptible to protease inhibitors.
Bacher
A
A
Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Garching, Germany.
Griebl
K
K
Mackamul
S
S
Mitreiter
R
R
Mückter
H
H
Ben-Shaul
Y
Y
D000976:Antipain / Q000494:pharmacology*; D005614:Freeze Fracturing; D006801:Humans; D007365:Intercellular Junctions / Q000378:metabolism* / Q000648:ultrastructure; D007976:Leupeptins / Q000494:pharmacology*; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism
D007976:Leupeptins; D011498:Protein Precursors; D000976:Antipain; C032854:leupeptin