ABC1, an ATP binding cassette transporter required for phagocytosis of apoptotic cells, generates a regulated anion flux after expression in Xenopus laevis oocytes.
The ATP binding cassette transporter ABC1 is a 220-kDa glycoprotein expressed by macrophages and required for engulfment of cells undergoing programmed cell death. Since members of this family of proteins such as P-glycoprotein and cystic fibrosis transmembrane conductance regulator share the ability to transport anions, we have investigated the transport capability of ABC1 expressed in Xenopus oocytes using iodide efflux and voltage-clamp techniques. We report here that ABC1 generates an anion flux sensitive to glibenclamide, sulfobromophthalein, and blockers of anion transporters. The anion flux generated by ABC1 is up-regulated by orthovanadate, cAMP, protein kinase A, and okadaic acid. In other ABC transporters, mutating the conserved lysine in the nucleotide binding folds was found to severely reduce or abolish hydrolysis of ATP, which in turn altered the activity of the transporter. In ABC1, replacement of the conserved lysine 1892 in the Walker A motif of the second nucleotide binding fold increased the basal ionic flux, did not alter the pharmacological inhibitory profile, but abolished the response to orthovanadate and cAMP agonists. Therefore, we conclude that ABC1 is a cAMP-dependent and sulfonylurea-sensitive anion transporter.
Becq
F
F
Laboratoire de Neurobiologie Cellulaire, CNRS, 31 Chemin J. Aiguier, 13402 Marseille Cedex 20, France.
Hamon
Y
Y
Bajetto
A
A
Gola
M
M
Verrier
B
B
Chimini
G
G
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters / Q000096:biosynthesis / Q000737:chemistry / Q000493:pharmacokinetics* / Q000502:physiology*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D000838:Anions / Q000378:metabolism; D017209:Apoptosis; D001483:Base Sequence; D017124:Conserved Sequence; D000242:Cyclic AMP / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000502:physiology; D005260:Female; D005905:Glyburide / Q000494:pharmacology; D006023:Glycoproteins / Q000096:biosynthesis / Q000737:chemistry / Q000493:pharmacokinetics*; D006801:Humans; D007454:Iodides / Q000378:metabolism*; D007700:Kinetics; D008239:Lysine; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D010587:Phagocytosis; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013448:Sulfobromophthalein / Q000494:pharmacology; D014158:Transcription, Genetic; D014638:Vanadates / Q000494:pharmacology; D014982:Xenopus laevis
C577836:ABCA1 protein, human; D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters; D000838:Anions; C505032:CFTR protein, human; D006023:Glycoproteins; D007454:Iodides; D011994:Recombinant Proteins; D013448:Sulfobromophthalein; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D014638:Vanadates; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D008239:Lysine; D005905:Glyburide; D015056:1-Methyl-3-isobutylxanthine