Band 3-mediated flip-flop and phosphatase-catalyzed cleavage of a long-chain alkyl phosphate anion in the human erythrocyte membrane.
In pursuit of the characterization of the recently discovered flippase mode of operation of the anion transporter (band 3, AE1) of the human erythrocyte membrane, the transbilayer translocation (flip) of a fluorescently labeled, membrane-intercalated long-chain alkyl phosphate, 10-(alpha-napthyl)-1-decyl-phosphate (NDP) was investigated. In contrast to the alkyl sulfonates and esters of phosphatidic acid studied as yet, NDP moves exclusively via band 3. NDP is, however, dephosphorylated at the inner membrane surface by a cytoplasmic phosphatase likely to interact specifically with endofacial membrane structures of the erythrocyte. This phosphatase shares characteristic inhibitor sensitivities with protein tyrosine phosphatases present in the erythrocyte interior. Vanadate as an inhibitor of NDP dephosphorylation provided a means to study the kinetic properties and patterns of inhibition (by inhibitors of anion exchange) and stimulation (by proteolysis of band 3 and aliphatic alcohols) of the flip of NDP. NDP is also an inhibitor of the exchange of hydrophilic anions via band 3, while hydrophilic anions interfere with the flip of NDP. The results are compared with the characteristics of the flip, via Band 3, of other amphiphilic anions and of the exchange of hydrophilic anions. Attempts are presented to understand the low flip rate of long-chain amphiphilic anions on the basis of their molecular properties and the thermodynamics of the "transition state" of the flip process.
Kleinhorst
A
A
Institut für Physiologie, Medizinische Fakultät der RWTH Aachen, Universitätsklinikum, Aachen, Germany.
Oslender
A
A
Haest
C W
CW
Deuticke
B
B
D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D004910:Erythrocyte Membrane / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D017136:Ion Transport; D008565:Membrane Proteins / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*
D001457:Anion Exchange Protein 1, Erythrocyte; D002352:Carrier Proteins; D008565:Membrane Proteins; D010710:Phosphates; D010744:Phosphoric Monoester Hydrolases