title: cGMP transport by vesicles from human and mouse erythrocytes.
abstract: cGMP secretion from cells can be mediated by ATP-binding cassette (ABC) transporters ABCC4, ABCC5, and ABCC11. Indirect evidence suggests that ABCC4 and ABCC5 contribute to cGMP transport by erythrocytes. We have re-investigated the issue using erythrocytes from wild-type and transporter knockout mice. Murine wild-type erythrocyte vesicles transported cGMP with an apparent Km that was 100-fold higher than their human counterparts, the apparent Vmax being similar. Whereas cGMP transport into human vesicles was efficiently inhibited by the ABCC4-specific substrate prostaglandin E1, cGMP transport into mouse vesicles was inhibited equally by Abcg2 and Abcc4 inhibitors/substrates. Similarly, cGMP transport into vesicles from Abcc4-/- and Abcg2-/- mice was 42% and 51% of that into wild-type mouse vesicles, respectively, whereas cGMP transport into vesicles from Abcc4(-/-)/Abcg2(-/-) mice was near background. The knockout mice were used to show that Abcg2-mediated cGMP transport occurred with lower affinity but higher Vmax than Abcc4-mediated transport. Involvement of Abcg2 in cGMP transport by Abcc4-/- erythrocyte vesicles was supported by higher transport at pH 5.5 than at pH 7.4, a characteristic of Abcg2-mediated transport. The relative contribution of ABCC4/Abcc4 and ABCG2/Abcg2 in cGMP transport was confirmed with a new inhibitor of ABCC4 transport, the protease inhibitor 4-(2-aminoethyl)benzenesulfonyl fluoride.
authors.affiliation: Department of Molecular Biology, the Netherlands Cancer Institute, Amsterdam, the Netherlands.
authors.forename: Cornelia J F
authors.initials: CJ
authors.lastname: de Wolf
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Yamaguchi
authors.forename: Ingrid
authors.initials: I
authors.lastname: van der Heijden
authors.forename: Peter R
authors.initials: PR
authors.lastname: Wielinga
authors.forename: Stefanie L
authors.initials: SL
authors.lastname: Hundscheid
authors.forename: Nobuhito
authors.initials: N
authors.lastname: Ono
authors.forename: George L
authors.initials: GL
authors.lastname: Scheffer
authors.forename: Marcel
authors.initials: M
authors.lastname: de Haas
authors.forename: John D
authors.initials: JD
authors.lastname: Schuetz
authors.forename: Jan
authors.initials: J
authors.lastname: Wijnholds
authors.forename: Piet
authors.initials: P
authors.lastname: Borst
mesh_terms: D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000737:chemistry / Q000235:genetics; D000527:Alprostadil / Q000378:metabolism; D000818:Animals; D001692:Biological Transport; D006152:Cyclic GMP / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004910:Erythrocyte Membrane / Q000378:metabolism; D004912:Erythrocytes / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics; D009363:Neoplasm Proteins / Q000235:genetics
chemical_list: D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D000527:Alprostadil; D006152:Cyclic GMP; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D009363:Neoplasm Proteins; C435500:ABCC11 protein, human; C073492:ABCC4 protein, human; C116614:ABCG2 protein, human; C489540:Abcc4 protein, mouse; C501412:Abcg2 protein, mouse