Protein-mediated chloride-phosphate and lactate-lactate exchange in cytoskeleton-free vesicles budded from rabbit erythrocytes.
Spectrin-free budded vesicles from rabbit erythrocytes (Leonards, K.S. and Ohki, S. (1983) Biochim. Biophys. Acta 728, 383-393) exchange intravesicular L-[14C]lactate for extravesicular L-lactate and intravesicular [36C]chloride for extravesicular phosphate with inhibitor sensitivity consistent with what is seen in intact cells. The time-course of these fluxes is faster than for intact cells, but is somewhat slower than predicted from surface to volume ratios. Labelling with tritiated 4,4'-diisothiocyanyl-2,2'-dihydrostilbenedisulfonate (H2DIDS) at concentrations which selectively inhibit inorganic anion exchange or specific lactate exchange supports the involvement of a 93-110 kDa (band 3) polypeptide in anion transport and a 40-50 kDa polypeptide in lactate transport across these vesicle membranes. Since the budded vesicles have a markedly simplified protein profile on electrophoresis, their isolated membranes represent a preliminary stage in the purification of these transport proteins in which structure and function appear to be preserved.
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000031:analogs & derivatives*; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid / Q000031:analogs & derivatives; D000345:Affinity Labels; D000818:Animals; D027321:Anion Transport Proteins; D001692:Biological Transport; D002352:Carrier Proteins / Q000097:blood; D002712:Chlorides / Q000097:blood*; D003598:Cytoskeletal Proteins / Q000097:blood; D004910:Erythrocyte Membrane / Q000378:metabolism*; D066298:In Vitro Techniques; D007773:Lactates / Q000097:blood*; D008565:Membrane Proteins / Q000097:blood / Q000502:physiology*; D008970:Molecular Weight; D027501:Monocarboxylic Acid Transporters; D010710:Phosphates / Q000097:blood*; D011817:Rabbits
D000345:Affinity Labels; D027321:Anion Transport Proteins; D002352:Carrier Proteins; D002712:Chlorides; D003598:Cytoskeletal Proteins; D007773:Lactates; D008565:Membrane Proteins; D027501:Monocarboxylic Acid Transporters; D010710:Phosphates; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; C018564:dihydro-DIDS; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid