Mediation of annexin 1 secretion by a probenecid-sensitive ABC-transporter in rat inflamed mucosa.
Annexin 1 is secreted by mammalian cells but lacks a leader signal sequence necessary to lead it to the classical secretory pathway via the endoplasmic reticulum. The mechanisms involved in the secretion of leaderless proteins remain uncertain. It has been suggested to involve membrane translocation via an ABC-transporter (ATP binding cassette). Using cultured inflamed mucosa from rectocolitis induced in rats, we studied if annexin 1 secretion followed the two main characteristics of ABC-transporter substrates: dependency on ATP hydrolysis and competitive inhibition by several other ABC-transporter substrates. Annexin 1 secretion is inhibited in a dose-dependent manner by two ATPase inhibitors. The inhibition reached 63.2+/-3.2%, 66.1+/-3.73% and 88.6+/-1.4% in the presence of 2mM vanadate, 0.5 and 1mM pervanadate, respectively. The efflux of calcein, a known ABC-transporter substrate, is similarly inhibited by 69.4+/-2.8% in the presence of 1mM pervanadate. Probenecid, an inhibitor of several ABC-transporters of the subfamilly ABCC or MRP (multidrug resistant associated protein), also inhibited annexin 1 secretion in a dose-dependent manner. As compared to control, 10mM probenecid reduced annexin 1 secretion by 72+/-20% and 20mM by 95.0+/-9%. By contrast, annexin 1 secretion is not blocked by other inhibitors of MRP1 (indomethacin, MK571), MRP2 (ochratoxin A1 or MK571), MRP5 (trequinsin or sulfinpyrazone) or by verapamil, cyclosporin A or glyburide. Taken together, our results show that annexin 1 secretion appears to share the efflux properties of ABC-transporter substrates.
Institut National de Recherche Agronomique, UR66, 180 Chemin de Tournefeuille, BP3, 31931 Toulouse Cedex 9, France.
Sharon
S
Wein
Myriam
M
Fauroux
Joëlle
J
Laffitte
Patricia
P
de Nadaï
Claire
C
Guaïni
François
F
Pons
Christine
C
Coméra
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D017305:Annexin A1 / Q000378:metabolism*; D001692:Biological Transport; D005452:Fluoresceins / Q000378:metabolism; D007249:Inflammation / Q000139:chemically induced / Q000473:pathology; D051379:Mice; D009092:Mucous Membrane / Q000378:metabolism*; D011339:Probenecid / Q000494:pharmacology; D051381:Rats; D014302:Trinitrobenzenesulfonic Acid
D018528:ATP-Binding Cassette Transporters; D017305:Annexin A1; D005452:Fluoresceins; D000255:Adenosine Triphosphate; D014302:Trinitrobenzenesulfonic Acid; D011339:Probenecid; C007740:fluorexon