Isolation and cDNA cloning of Ksp-cadherin, a novel kidney-specific member of the cadherin multigene family.
AuthorsThomson R B; Igarashi P; Biemesderfer D; Kim R; Abu-Alfa A; Soleimani M; Aronson P S;
JournalJ Biol Chem
PubMed ID7615566
Cadherins are recognized as the principal mediators of homotypic cellular recognition and play a demonstrated role in the morphogenic direction of tissue development. We report here the identification of a structurally unique, kidney-specific member of the cadherin multigene family (Ksp-cadherin). cDNA cloning and molecular analysis of the 130-kDa protein confirmed ... More
The mechanisms of inhibition of anion exchange in human erythrocytes by 1-ethyl-3-[3-(trimethylammonio)propyl]carbodiimide.
AuthorsWerner PK, Reithmeier RA
JournalBiochim Biophys Acta
PubMed ID2454665
'Treatment of human erythrocytes with the membrane-impermeant carbodiimide 1-ethyl-3-[3-(trimethylammonio)propyl]carbodiimide (ETC) in citrate-buffered sucrose leads to irreversible inhibition of phosphate-chloride exchange. The level of transport inhibition produced was dependent on the concentration of citrate present during treatment, with a maximum of approx. 60% inhibition. [14C]Citric acid was incorporated into Band 3 ... More
The effect of apical and basolateral lipids on the function of the band 3 anion exchange protein.
Authorsvan't Hof W, Malik A, Vijayakumar S, Qiao J, van Adelsberg J, Al-Awqati Q
JournalJ Cell Biol
PubMed ID9362512
'Although many polarized proteins are sorted to the same membrane domain in all epithelial tissues, there are some that exhibit a cell type-specific polarity. We recently found that band 3 (the anion exchanger AE1) was present in the apical membrane of a renal intercalated cell line when these cells were ... More
35Cl nuclear magnetic resonance line broadening shows that eosin-5-maleimide does not block the external anion access channel of band 3.
AuthorsLiu D, Kennedy SD, Knauf PA
JournalBiophys J
PubMed ID8527653
'It has been suggested that Lys-430 of band 3, with which eosin-5-maleimide (EM) reacts, is located in the external channel through which anions gain access to the external transport site, and that EM inhibits anion exchange by blocking this channel. To test this, we have used 35Cl nuclear magnetic resonance ... More
Identification and partial purification of the erythrocyte L-lactate transporter.
AuthorsPoole RC, Halestrap AP
JournalBiochem J
PubMed ID1590773
'1. Intact erythrocytes were incubated with 100 microM-4,4''-di-isothiocyanostilbene-2,2''-disulphonate (DIDS), a concentration sufficient to inhibit lactate transport irreversibly by 65%. DIDS-labelled proteins were detected by immunoblotting of erythrocyte membrane proteins with an anti-DIDS antibody. Labelled polypeptides of 35-45 kDa in rat erythrocytes, and of 40-50 kDa in rabbit and guinea pig ... More
Reversible and irreversible inhibition, by stilbenedisulphonates, of lactate transport into rat erythrocytes. Identification of some new high-affinity inhibitors.
AuthorsPoole RC, Halestrap AP
JournalBiochem J
PubMed ID2025218
'1. Inhibition of L-lactate transport into rat erythrocytes by stilbenedisulphonates was studied under conditions which allowed the contribution of reversible and irreversible inhibition to be assessed. 2. At low temperatures (7 degrees C), 4,4''-di-isothiocyanostilbene-2,2''-disulphonate (DIDS) and other stilbenedisulphonates were found to inhibit lactate transport instantaneously, in a manner which was ... More
Location of the stilbenedisulfonate binding site of the human erythrocyte anion-exchange system by resonance energy transfer.
AuthorsRao A, Martin P, Reithmeier RA, Cantley LC
JournalBiochemistry
PubMed ID497152
'The stilbenedisulfonate inhibitory site of the human erythrocyte anion-exchange system has been characterized by using serveral fluorescent stilbenedisulfonates. The covalent inhibitor 4-benzamido-4''-isothiocyanostilbene-2,2''-disulfonate (BIDS) reacts specifically with the band 3 protein of the plasma membrane when added to intact erythrocytes, and the reversible inhibitors 4,4''-dibenzamidostilbene-2,2''-disulfonate (DBDS) and 4-benzamido-4''-aminostilbene-2,2''-disulfonate (BADS) show a ... More
Trapping of inhibitor-induced conformational changes in the erythrocyte membrane anion exchanger AE1.
AuthorsTang XB, Casey JR
JournalBiochemistry
PubMed ID10545179
'AE1, the chloride/bicarbonate anion exchanger of the erythrocyte plasma membrane, is highly sensitive to inhibition by stilbene disulfonate compounds such as DIDS (4,4''-diisothiocyanostilbene-2, 2''-disulfonate) and DNDS (4,4''-dinitrostilbene-2,2''-disulfonate). Stilbene disulfonates recruit the anion binding site to an outward-facing conformation. We sought to identify the regions of AE1 that undergo conformational changes ... More
Intracellular pH-regulating mechanism of the squid axon. Interaction between DNDS and extracellular Na+ and HCO3-.
AuthorsBoron WF, Knakal RC
JournalJ Gen Physiol
PubMed ID2915212
'Intracellular pH (pHi) of the squid axon is regulated by a stilbenesensitive transporter that couples the influx of Na+ and HCO3- (or the equivalent) to the efflux of Cl-. According to one model, the extracellular ion pair NaCO3- exchanges for intracellular Cl-. In the present study, the ion-pair model was ... More
Characterization of the stilbenedisulfonate binding site on band 3.
AuthorsSchopfer LM, Salhany JM
JournalBiochemistry
PubMed ID7599124
'Stilbenedisulfonates are potent inhibitors of Band 3 mediated anion exchange. They bind tightly to the protein and form a 1-to-1 reversible complex. Those stilbenedisulfonates which contain isothocyanato groups such as DIDS (4,4'-diisothiocyanato-2,2'-stilbenedisulfonate) and H2DIDS (4,4'-diisothiocyanatodihydrostilbene-2,2'-disulfonate) can also react rapidly with lysine residues within the binding pocket to yield an irreversible ... More
Characterization of the osteoclast ruffled border chloride channel and its role in bone resorption.
'Bone resorption by osteoclasts requires massive transcellular acid transport, which is accomplished by the parallel action of a V-type proton pump and a chloride channel in the osteoclast ruffled border. We have studied the molecular basis for the appearance of acid transport as avian bone marrow mononuclear cells acquire a ... More
Interactions of inhibitors on anion transporter of human erythrocyte.
AuthorsFröhlich O, Gunn RB
JournalAm J Physiol
PubMed ID3826335
'Chloride tracer efflux was measured from intact human erythrocytes into media containing different chloride concentrations and different concentrations of the inhibitors 4,4''-dinitrostilbene-2-2''-disulfonate (DNDS), N-(4-azido-2-nitrophenyl)-2-aminoethylsulfonate (NAP-taurine), phloretin, and sulfate. The data were analyzed to test whether these inhibitors were mutually exclusive with each other or whether they could bind at the ... More
Effects of inhibitors on anion exchangers in rabbit renal brush border membrane vesicles.
AuthorsMcConnell KR, Aronson PS
JournalJ Biol Chem
PubMed ID8063783
'Cl-formate, Cl-oxalate, and SO4-CO3 exchange participate in Cl and organic anion transport across the brush border membrane of the rabbit proximal tubule. To determine the functional similarity of these transporters to each other and to band 3, we characterized, in isolated membrane vesicles, the inhibition of these transporters by compounds ... More
Kinetic evidence for ternary complex formation and allosteric interactions in chloride and stilbenedisulfonate binding to band 3.
AuthorsSalhany JM, Sloan RL, Cordes KA, Schopfer LM
JournalBiochemistry
PubMed ID7918409
'The molecular basis for chloride and stilbenedisulfonate interaction with band 3 was investigated by measuring the kinetics of stilbenedisulfonate release from its complex with the transporter. We found that 150 mM NaCl accelerated the rate of release of DBDS (4,4''-dibenzamidostilbene-2,2''-dibenzamidostilbene-2,2''-disu lfonate) and H2DIDS (4,4''-diisothiocyanodihydrostilbene-2,2''-disulfonate) by more than 10-fold at constant ... More
Chloride channel inhibition by the venom of the scorpion Leiurus quinquestriatus.
AuthorsDeBin JA, Strichartz GR
JournalToxicon
PubMed ID1726031
'The venom of the scorpion Leiurus quinquestriatus produced a significant, reversible inhibition of reconstituted Cl- channels of the small conductance type found in rat colonic epithelial cells. The kinetics of single-channel block by this venom were consistent with a first-order binding reaction in which the binding of one ligand molecule ... More
Endotoxin-induced degranulation of the Limulus amebocyte.
AuthorsArmstrong PB, Rickles FR
JournalExp Cell Res
PubMed ID6286331
Electron spin resonance studies on the inorganic-anion-transport system of the human red blood cell. Binding of a disulfonatostilbene spin label (NDS-TEMPO) and inhibition of anion transport.
AuthorsSchnell KF, Elbe W, Käsbauer J, Kaufmann E
JournalBiochim Biophys Acta
PubMed ID6307363
'The disulfonatostilbene spin label, NDS-TEMPO, was synthesized (purity over 96%) and the binding of the spin label to human red-cell ghosts was studied. NDS-TEMPO is readily absorbed to the membrane surface. Both pretreatment of the ghosts with FDNB and DIDS and the presence of DNDS completely prevent the binding of ... More
Roles of histidine 752 and glutamate 699 in the pH dependence of mouse band 3 protein-mediated anion transport.
AuthorsMüller-Berger S, Karbach D, Kang D, Aranibar N, Wood PG, Rüterjans H, Passow H
JournalBiochemistry
PubMed ID7626601
'In the accompanying paper we have shown that four different histidine residues are involved in the maintenance of mouse band 3 in a state in which it is able to execute its anion transport function. Here we focus on the functional significance of His 752 and demonstrate that this residue, ... More
Self-association of Band 3, the human erythrocyte anion exchanger, in detergent solution.
AuthorsVince JW, Sarabia VE, Reithmeier RA
JournalBiochim Biophys Acta
PubMed ID9218560
'Dimeric Band 3 purified in n-dodecyl octaethyleneglycol (C12E8) underwent an irreversible, temperature-dependent association, resulting in a complex with a Stokes radius slightly larger than a native tetramer, before forming a higher molecular weight aggregate. Self-association occurred with a half-time of about 1 h at 37 degrees C but did not ... More
Fluorescence labeling of the human erythrocyte anion transport system.
AuthorsDissing S, Jesaitis AJ, Fortes PA
JournalBiochim Biophys Acta
PubMed ID454588
'The anion transport system of human red cells was isolated in vesicles containing the original membrane lipids and the 95 000 dalton polypeptides (band 3) by the method of Wolosin et al. (J. Biol. Chem. (1977) 252, 2419--2427). The vesicles have a functional anion transprot system since they display sulfate ... More
Bicarbonate-dependent chloride secretion in Calu-3 epithelia in response to 7,8-benzoquinoline.
AuthorsCuthbert AW, Supuran CT, MacVinish LJ
JournalJ Physiol
PubMed ID12872009
'Stimulation of Calu-3 epithelia with 7,8-benzoquinoline, under short circuit current conditions, produced a current increase that was completely accounted for by the net flux of chloride, measured simultaneously with 36Cl-. Nevertheless the current stimulated by 7,8-benzoquinoline was sensitive to acetazolamide, which caused up to 50 % inhibition of the stimulated ... More
Effects of pH on kinetic parameters of the Na-HCO3 cotransporter in renal proximal tubule.
AuthorsGross E, Hopfer U
JournalBiophys J
PubMed ID10354432
'The effects of pH on cotransporter kinetics were studied in renal proximal tubule cells. Cells were grown to confluence on permeable support, mounted in an Ussing-type chamber, and permeabilized apically to small monovalent ions with amphotericin B. The steady-state, dinitrostilbene-disulfonate-sensitive current (DeltaI) was Na+ and HCO3- dependent and therefore was ... More
Purification and characterization of chlorotoxin, a chloride channel ligand from the venom of the scorpion.
AuthorsDeBin JA, Maggio JE, Strichartz GR
JournalAm J Physiol
PubMed ID8383429
We have previously demonstrated that the venom of the scorpion Leiurus quinquestriatus blocks small-conductance Cl- channels, derived from epithelial cells, when applied to the cytoplasmic surface. We have now purified to near homogeneity, and characterized, the component responsible for this blocking activity. It is a small basic peptide of 4,070 ... More
Band 3, an accidental flippase for anionic phospholipids?
AuthorsVondenhof A, Oslender A, Deuticke B, Haest CW
JournalBiochemistry
PubMed ID8161506
The inward translocation of the monovalent anionic phospholipid 1-palmitoyl-sn-glycero-3-phosphomethanol in the membrane of human erythrocytes is a fast process (t/2 = 11 min, 37 degrees C). Translocation of the protonated uncharged phospholipid is not responsible for the fast flip rate, and mediation of translocation by the aminophospholipid flippase could be ... More
Functional evidence for distinct interaction of hydrophobic arylisothiocyanates with the erythrocyte anion transport protein.
AuthorsCacciola SO, Sigrist H, Reist M, Cabantchik ZI, Zahler P
JournalJ Membr Biol
PubMed ID6492134
Human erythrocytes were treated with various hydrophobic arylisothiocyanates under conditions which favor modification of distinct proteinaceous nucleophiles. The morphological appearance of phenylisothiocyanate-treated cells was discoid and membrane-bound hydrolases (human acetylcholinesterase, sheep phospholipase A2) were fully active following membrane modification. Noncharged hydrophobic arylisothiocyanates, including phenylisothiocyanate, beta-naphthylisothiocyanate and heterobifunctional azidoarylisothiocyanates inhibited [35S]-sulfate ... More
Protein kinase A regulates chloride conductance in endocytic vesicles from proximal tubule.
AuthorsBae HR, Verkman AS
JournalNature
PubMed ID1701220
Regulation of ion transport by phosphorylation and G proteins occurs in several epithelial and non-epithelial cell plasma membranes1-5. It is not known whether transporters on intracellular membranes are target sites for second messengers. Here we present direct evidence that a chloride conductance in endocytic vesicles from rabbit proximal tubule is ... More
Searching for mechanisms of N-methyl-D-aspartate-induced glutathione efflux in organotypic hippocampal cultures.
AuthorsWallin C, Abbas AK, Tranberg M, Weber SG, Wigström H, Sandberg M
JournalNeurochem Res
PubMed ID12608701
N-Methyl-D-aspartate (NMDA)-receptor stimulation evoked a selective and partly delayed elevated efflux of glutathione, phosphoethanolamine, and taurine from organotypic rat hippocampus slice cultures. The protein kinase inhibitors H9 and staurosporine had no effect on the efflux. The phospholipase A2 inhibitors quinacrine and 4-bromophenacyl bromide, as well as arachidonic acid, a product ... More
Inhibition of anion transport in the human red blood cell membrane with para- and meta-methoxyphenylglyoxal.
AuthorsBetakis E, Fritzsch G, Zaki L
JournalBiochim Biophys Acta
PubMed ID1390838
The positional isomers para-methoxyphenylglyoxal and meta-methoxyphenylglyoxal were newly synthesized and found to be potent inhibitors of sulfate exchange in the red blood cell membrane. The rate of inactivation of the transport system with both reagents obeys pseudo-first-order kinetics and increases with increasing pH and reagent concentration. The degree of inhibition ... More
Chloride channel blockers inhibit Ca2+ uptake by the smooth muscle sarcoplasmic reticulum.
AuthorsPollock NS, Kargacin ME, Kargacin GJ
JournalBiophys J
PubMed ID9746517
Despite the fact that Ca2+ transport into the sarcoplasmic reticulum (SR) of muscle cells is electrogenic, a potential difference is not maintained across the SR membrane. To achieve electroneutrality, compensatory charge movement must occur during Ca2+ uptake. To examine the role of Cl- in this charge movement in smooth muscle ... More
Complementation studies with Co-expressed fragments of the human red cell anion transporter (Band 3; AE1). The role of some exofacial loops in anion transport.
AuthorsWang L, Groves JD, Mawby WJ, Tanner MJ
JournalJ Biol Chem
PubMed ID9099711
We constructed cDNA clones encoding fragments of band 3 in which the membrane domain was truncated from either the N or the C terminus within each of the first four exofacial loops. The truncations containing the C terminus of the protein were fused with the cleavable N-terminal signal sequence of ... More
Histidine-834 of human erythrocyte band 3 has an essential role in the conformational changes that occur during the band 3-mediated anion exchange.
AuthorsJin XR, Abe Y, Li CY, Hamasaki N
JournalBiochemistry
PubMed ID14596607
We have shown that diethyl pyrocarbonate (DEPC) inhibits band 3-mediated anion exchange and that the inhibition occurs only when histidine residue(s) is (are) modified with DEPC from the cytosolic surface of resealed ghosts [Izuhara et al. (1989) Biochemistry 28, 4725-4728]. In the present study, we have identified the DEPC-modified histidine ... More
Anion transport in red blood cells. III. Sites and sidedness of inhibition by high-affinity reversible binding probes.
AuthorsBarzilay M, Cabantchik ZI
JournalMembr Biochem
PubMed ID514089
Studies of binding of the reversible inhibitor DNDS (for abbreviations, see Nomenclature) and red blood cell membranes revealed 8.6 +/- 0.7 x 10(5) high-affinity binding sites per cell (KD = 0.8 +/- 0.4 muM). Under conditions of "mutual depletion," inhibition studies of anion exchange revealed 8.0 +/- 0.7 x 10(5) ... More
Macromolecular conjugates of transport inhibitors: new tools for probing topography of anion transport proteins.
AuthorsEidelman O, Yani P, Englert HC, Lang HG, Greger R, Cabantchik ZI
JournalAm J Physiol
PubMed ID2035616
Macromolecular-conjugated, water-soluble, membrane-impermeant compounds were designed and assessed as topological probes for chloride-transporting agencies. The novel compounds were derivatives of either disulfonic stilbene (DS) and benzylaminoethylsulfonate (BS), "classical" inhibitors of erythrocyte chloride-bicarbonate exchange, or of phenylanthranilates (PA), high-affinity blockers of epithelial chloride channels. Covalent reactive derivatives of various DS, BS, ... More
Protein kinase A dependent membrane protein phosphorylation and chloride conductance in endosomal vesicles from kidney cortex.
AuthorsReenstra WW, Sabolic I, Bae HR, Verkman AS
JournalBiochemistry
PubMed ID1310027
Regulation of Cl conductance by protein kinase A may play a role in control of endosomal acidification [Bae, H.-R., & Verkman, A. S. (1990) Nature, 348, 637-639]. To investigate the mechanism of kinase A action, cell-free measurements of Cl transport and membrane protein phosphorylation were carried out in apical endocytic ... More
Detection of Cl- binding to band 3 by double-quantum-filtered 35Cl nuclear magnetic resonance.
AuthorsLiu D, Knauf PA, Kennedy SD
JournalBiophys J
PubMed ID8789088
We have applied double-quantum-filtered (DQF) NMR of 35Cl to study binding of Cl- to external sites on intact red blood cells, including the outward-facing anion transport sites of band 3, an integral membrane protein. A DQF 35Cl NMR signal was observed in cell suspensions containing 150 mM KCl, but the ... More
Voltage and cosubstrate dependence of the Na-HCO3 cotransporter kinetics in renal proximal tubule cells.
AuthorsGross E, Hopfer U
JournalBiophys J
PubMed ID9675182
The voltage dependence of the kinetics of the sodium bicarbonate cotransporter was studied in proximal tubule cells. This electrogenic cotransporter transports one Na+, three HCO3-, and two negative charges. Cells were grown to confluence on a permeable support, mounted on a Ussing-type chamber, and permeabilized apically to small monovalent ions ... More
Molecular mechanisms of band 3 inhibitors. 1. Transport site inhibitors.
AuthorsFalke JJ, Chan SI
JournalBiochemistry
PubMed ID3801446
The band 3 protein of red cells is a transmembrane ion transport protein that catalyzes the one-for-one exchange of anions across the cell membrane. 35Cl NMR studies of Cl- binding to the transport sites of band 3 show that inhibitors of anion transport can be grouped into three classes: (1) ... More
Preliminary pharmacological characterisation of an interleukin-13-enhanced calcium-activated chloride conductance in the human airway epithelium.
AuthorsAtherton H, Mesher J, Poll CT, Danahay H
JournalNaunyn Schmiedebergs Arch Pharmacol
PubMed ID12595965
Interleukin (IL)-13 (10 ng/ml for 48 h) treatment of human bronchial epithelial cells induced a hypersecretory ion transport phenotype. Ussing chamber experiments demonstrated that this phenotypic change was characterised by an almost complete inhibition of the amiloride-sensitive short circuit current (ISC) and the appearance of an enhanced calcium-activated chloride conductance ... More
Band-3 mediated uptake of beryllofluoride complexes by human erythrocytes.
AuthorsXu AS, Morris MB, Kuchel PW
JournalBiochemistry
PubMed ID1390712
Beryllium forms several multivalent fluoride complexes in aqueous solution; the relative concentration of each is governed by the relative concentrations of the constituent ions and pH. In 9Be NMR spectra the 9Be (spin = 3/2) and 19F (spin = 1/2) spin coupling gave rise to an overlapping resonance triplet, quartet, ... More
An oxonol dye is the most potent known inhibitor of band 3-mediated anion exchange.
AuthorsKnauf PA, Law FY, Hahn K
JournalAm J Physiol
PubMed ID7485448
When cells are acutely exposed to the oxonol dye, bis(1,3-dibutylbarbituric acid)pentamethine oxonol (diBA), at 0 degrees C, the concentration that gives half inhibition of Cl- exchange (IC50) is 0.146 +/- 0.013 microM (n = 12) initially, but the inhibition increases with time. These characteristics indicate that a rapid initial binding ... More
High affinity binding of ankyrin induced by volume expansion in skate erythrocytes.
AuthorsMusch MW, Goldstein L
JournalJ Biol Chem
PubMed ID8702894
Volume expansion of little skate (Raja erinacea) erythrocytes increases the affinity of ankyrin binding without altering in the number of binding sites. Potassium iodide-stripped inside-out vesicles (KI-IOV) were used to assess ankyrin binding under volume-expanded conditions. Under isoosmotic conditions, ankyrin binds nearly exclusively to a single class of sites (Bmax, ... More
Chemical probes for anion transporters of mammalian cell membranes.
AuthorsCabantchik ZI, Greger R
JournalAm J Physiol
PubMed ID1566811
Mammalian cell membranes harbor several types of chloride channels, chloride-cation symporters/cotransporters, and several classes of anion exchangers/antiporters. These transport systems subserve different cellular or organismic functions, depending on the nature of the cell, the spatial organization of transporters, and their functional interplay. Chemical probing has played a central role in ... More
Inhibition and derivatization of the renal Na,K-ATPase by dihydro-4,4'-diisothiocyanatostilbene-2,2'-disulfonate.
AuthorsPedemonte CH, Kaplan JH
JournalBiochemistry
PubMed ID2849997
Treatment of purified renal Na,K-ATPase with dihydro-4,4'-diisothiocyanatostilbene-2,2'-disulfonate (H2DIDS) produces both reversible and irreversible inhibition of the enzyme activity. The reversible inhibition is unaffected by the presence of saturating concentrations of the sodium pump ligands Na+,K+, Mg2+, and ATP, while the inactivation is prevented by either ATP or K+. The kinetics ... More
Murine T lymphocytes modulate activity of an ATP-activated P2Z-type purinoceptor during differentiation.
AuthorsChused TM, Apasov S, Sitkovsky M
JournalJ Immunol
PubMed ID8759716
Murine T, but not B, lymphocytes constitutively express a membrane receptor for adenosine nucleotides that opens a nonspecific pore that admits Ca2+ and ethidium (314 Da), but not propidium (415 Da) ions. ATP, ADP, and AMP show decreasing potency; UTP and adenosine are inactive. Nonhydrolyzable ATP analogues are completely ineffective. ... More
Interaction of the erythrocyte lactate transporter (monocarboxylate transporter 1) with an integral 70-kDa membrane glycoprotein of the immunoglobulin superfamily.
AuthorsPoole RC, Halestrap AP
JournalJ Biol Chem
PubMed ID9169423
Treatment of intact erythrocytes with 4,4'-diisothiocyanostilbene-2, 2'-disulfonate (DIDS) causes irreversible inhibition and chemical labeling of the lactate transporter, monocarboxylate transporter 1 (MCT1) (Poole, R. C., and Halestrap, A. P. (1992) Biochem. J. 283, 855-862). In rat erythrocytes DIDS also causes cross-linking of MCT1 to another protein in the membrane to ... More
High level expression, partial purification, and functional reconstitution of the human AE1 anion exchanger in Saccharomyces cerevisiae.
AuthorsSekler I, Kopito R, Casey JR
JournalJ Biol Chem
PubMed ID7673129
Human erythroid anion exchanger AE1 (Band 3) was expressed in the yeast Saccharomyces cerevisiae under the control of the constitutive promoter and transcriptional terminator of the yeast phosphoglycerate kinase gene. AE1 expression in stable yeast transformants was estimated to be approximately 0.7 mg AE1 per liter. Density gradient sedimentation analysis ... More
The external anion binding site of the human erythrocyte anion transporter: DNDS binding and competition with chloride.
AuthorsFröhlich O
JournalJ Membr Biol
PubMed ID7057455
The interaction between chloride and the anion transport inhibitor DNDS (4,4'-dinitro stilbene-2,2'-disulfonate) at the external anion binding site of the human erythrocyte anion transporter was examined by two techniques: a) chloride tracer flux experiments in the presence of varying concentrations of DNDS, and b) DNDS equilibrium binding experiments in the ... More
Airway surface liquid pH in well-differentiated airway epithelial cell cultures and mouse trachea.
AuthorsJayaraman S, Song Y, Verkman AS
JournalAm J Physiol Cell Physiol
PubMed ID11600413
Airway surface liquid (ASL) pH has been proposed to be important in the pathophysiology of cystic fibrosis, asthma, and cough. Ratio image analysis was used to measure pH in the ASL after staining with the fluorescent pH indicator 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF)-dextran. ASL pH in bovine airway cell cultures grown at an ... More
Sulfate transport mediated by the mammalian anion exchangers in reconstituted proteoliposomes.
AuthorsSekler I, Lo RS, Mastrocola T, Kopito RR
JournalJ Biol Chem
PubMed ID7744759
The kinetic properties of sulfate transport mediated by the anion exchangers AE1 and AE2 have been examined. Microsomes isolated from HEK cells transiently overexpressing either protein were reconstituted in unilamellar, 200-600-nm diameter proteoliposomes. Transport mediated by the exchangers was monitored by loading the reconstituted proteoliposomes with the slowly transportable anion ... More
Regulation of the calcium release channel from skeletal muscle by suramin and the disulfonated stilbene derivatives DIDS, DBDS, and DNDS.
AuthorsO'Neill ER, Sakowska MM, Laver DR
JournalBiophys J
PubMed ID12609870
Activation of skeletal muscle ryanodine receptors (RyRs) by suramin and disulfonic stilbene derivatives (Diisothiocyanostilbene-2',2'-disulfonic acid (DIDS), 4,4'-dibenzamidostilbene-2,2'-disulfonic acid (DBDS),and 4,4'-dinitrostilbene-2,2'-disulfonic acid (DNDS)) was investigated using planar bilayers. One reversible and two nonreversible mechanisms were identified. K(a) for reversible activation (approximately 100 micro M) depended on cytoplasmic [Ca(2+)] and the bilayer ... More
Inhibition of inorganic anion transport across the human red blood cell membrane by chloride-dependent association of dipyridamole with a stilbene disulfonate binding site on the band 3 protein.
AuthorsLegrum B, Passow H
JournalBiochim Biophys Acta
PubMed ID2923878
The inhibition of inorganic anion transport by dipyridamole (2,6-bis(diethanolamino)-4,8-dipiperidinopyrimido[5,4-d] pyrimidine) takes place only in the presence of Cl-, other halides, nitrate or bicarbonate. At any given dipyridamole concentration, the anion flux relative to the flux in the absence of dipyridamole follows the equation: Jrel = (1 + alpha 2[Cl-])/(1 + ... More
New approaches for the reconstitution and functional assay of membrane transport proteins. Application to the anion transporter of human erythrocytes.
AuthorsDarmon A, Zangvill M, Cabantchik ZI
JournalBiochim Biophys Acta
PubMed ID6824657
The human red blood cell anion transport protein, band 3, was isolated and reconstituted into lipid vesicles. The main feature of the new reconstitution is the replacement of native lipids and of solubilizing detergent by externally added lipids, while band 3 protein is immobilized on a gel matrix. The vesicles ... More
Anion transport in red blood cells. II. Kinetics of reversible inhibition by nitroaromatic sulfonic acids.
AuthorsBarzilay M, Cabantchik ZI
JournalMembr Biochem
PubMed ID229385
The anion exchange system of human red blood cells is highly inhibited and specifically labeled by isothiocyano derivatives of benzene sulfonate (BS) or stilbene disulfonate (DS). To learn about the site of action of these irreversibly binding probes we studied the mechanism of inhibition of anion exchange by the reversibly ... More
Inhibition of mouse erythroid band 3-mediated chloride transport by site-directed mutagenesis of histidine residues and its reversal by second site mutation of Lys 558, the locus of covalent H2DIDS binding.
AuthorsMüller-Berger S, Karbach D, König J, Lepke S, Wood PG, Appelhans H, Passow H
JournalBiochemistry
PubMed ID7626600
Substitution by site-directed mutagenesis of any one of the histidine residues H721, H837, and H852 by glutamine, or of H752 by serine, inhibits Cl- flux mediated by band 3 expressed in Xenopus oocytes. Mutation of Lys 558 (K558N), the site of covalent binding of H2DIDS (4,4'-diisothiocyanostilbene-2,2'-disulfonate) in the outer membrane ... More
The kinetics, substrate and inhibitor specificity of the lactate transporter of Ehrlich-Lettre tumour cells studied with the intracellular pH indicator BCECF.
AuthorsCarpenter L, Halestrap AP
JournalBiochem J
PubMed ID7818477
1. Suspensions of cultured Ehrlich-Lettre tumour cells were loaded with the pH-sensitive fluorescent indicator 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF), and changes in intracellular pH upon addition of L-lactate and other monocarboxylates were continuously monitored by fluorimetry using dual-wavelength excitation (450/500 nm) and single-wavelength emission (> 520 nm). 2. The rapid fluorescence changes were ... More
pH-dependence of inhibition by H2DIDS of mouse erythroid band 3-mediated Cl- transport in Xenopus oocytes. The effect of oligonucleotide-directed replacement of Lys-558 by an Asn residue.
AuthorsKietz D, Bartel D, Lepke S, Passow H
JournalBiochim Biophys Acta
PubMed ID1902748
The rapid reversible inhibition of band 3-mediated inorganic anion transport by 4,4'-diisothiocyanodihydrostilbene-2,2'-disfulfonate (H2DIDS) turns slowly into irreversible inhibition. This is due to covalent bond formation of the two isothiocyanate groups of the inhibitor with two lysine residues on band 3, called Lys a and Lys b. In the red cell ... More
The kinetics, substrate, and inhibitor specificity of the monocarboxylate (lactate) transporter of rat liver cells determined using the fluorescent intracellular pH indicator, 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein.
AuthorsJackson VN, Halestrap AP
JournalJ Biol Chem
PubMed ID8557697
The kinetics of transport of L-lactate, pyruvate, ketone bodies, and other monocarboxylates into isolated hepatocytes from starved rats were measured at 25 degrees C using the intracellular pH-sensitive dye, 2',7'-bis(carboxyethyl)- 5(6)-carboxyfluorescein, to detect the associated proton influx. Transport kinetics were similar, but not identical, to those determined using the same ... More
AuthorsFurukawa T, Virág L, Sawanobori T, Hiraoka M
JournalJ Membr Biol
PubMed ID8114079
Effects of stilbene disulfonates on single KATP channel currents were investigated in inside-out and outside-out membrane patches from guinea pig ventricular myocytes. All drugs tested, 4,4'-diisothiocyanatostilbene,2,2'-disulfonic acid (DIDS), 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid (SITS), 4,4'-dinitrostilbene-2,2'-disulfonic acid (DNDS), and 4,4'-diaminostilbene-2,2'-disulfonic acid (DADS), inhibited the KATP channel when they were applied to the intracellular, but ... More
n-Alkanols and halothane inhibit red cell anion transport and increase band 3 conformational change rate.
AuthorsForman SA, Verkman AS, Dix JA, Solomon AK
JournalBiochemistry
PubMed ID4074663
The effects of halothane and n-alkanols on band 3, the anion-exchange protein of the red cell membrane, have been characterized by radioactive sulfate exchange and equilibrium and kinetic binding of a fluorescent anion transport inhibitor, 4,4'-dibenzamido-2,2'-stilbenedisulfonic acid (DBDS), with fluorescence and stopped-flow techniques. Ethanol, butanol, hexanol, heptanol, octanol, and decanol ... More
Functional carboxyl groups in the red cell anion exchange protein. Modification with an impermeant carbodiimide.
AuthorsBjerrum PJ, Andersen OS, Borders CL, Wieth JO
JournalJ Gen Physiol
PubMed ID2738575
Anion exchange in human red blood cell membranes was inactivated using the impermeant carbodiimide 1-ethyl-3-(4-azonia-4,4-dimethylpentyl)-carbodiimide (EAC). The inactivation time course was biphasic: at 30 mM EAC, approximately 50% of the exchange capacity was inactivated within approximately 15 min; this was followed by a phase in which irreversible exchange inactivation was ... More
The kinetics of intramolecular cross-linking of the band 3 protein in the red blood cell membrane by 4,4'-diisothiocyano dihydrostilbene-2,2'-disulfonic acid (H2DIDS).
AuthorsKampmann L, Lepke S, Fasold H, Fritzsch G, Passow H
JournalJ Membr Biol
PubMed ID7186941
The two isothiocyanate groups of the anion transport inhibitor 4,4'-diisothiocyano dihydrostilbene-2-2'-disulfonate (H2DIDS) may react covalently with two lysine residues called a and b that reside on the chymotryptic 60,000 Dalton and 35,000 Dalton segments, respectively, of the band 3 protein of the human erythrocyte membrane. Under suitable conditions, the reaction ... More
Phosphorylation-induced modulation of pNBC1 function: distinct roles for the amino- and carboxy-termini.
AuthorsGross E, Fedotoff O, Pushkin A, Abuladze N, Newman D, Kurtz I
JournalJ Physiol
PubMed ID12730338
The human NBC1 (SLC4A4) gene encodes the electrogenic sodium bicarbonate cotransporters kNBC1 and pNBC1, which are highly expressed in the kidney and pancreas, respectively. The HCO3-:Na+ stoichiometry of these cotransporters is an important determinant of the direction of ion flux. Recently we showed in a mouse proximal tubule (mPCT) cell ... More
Oligomeric forms of skate erythrocyte band 3. Effect of volume expansion.
AuthorsMusch MW, Davis EM, Goldstein L
JournalJ Biol Chem
PubMed ID8051044
Volume expansion of little skate (Raja erinacea) erythrocytes leads to the formation of oligomeric forms of the band 3 protein. The oligomers are formed following incubation in hypotonic media or inclusion of a permeant solute such as ethylene glycol. Oligomers were detected by specifically labeling the skate erythrocyte band 3 ... More
Regulation of intracellular pH in Calu-3 human airway cells.
AuthorsInglis SK, Finlay L, Ramminger SJ, Richard K, Ward MR, Wilson SM, Olver RE
JournalJ Physiol
PubMed ID11790817
The Calu-3 human cell line exhibits features of submucosal gland serous cells and secretes HCO(3)(-). The aim of this study was to identify the HCO(3)(-) transporters present in these cells by studying their role in the regulation of intracellular pH (pH(i)). Calu-3 cells were grown on coverslips, loaded with the ... More
Inhibition and stimulation of phospholipid scrambling activity. Consequences for lipid asymmetry, echinocytosis, and microvesiculation of erythrocytes.
AuthorsKamp D, Sieberg T, Haest CW
JournalBiochemistry
PubMed ID11478914
An increase of the intracellular Ca(2+) concentration in erythrocytes is known to activate rapid nonspecific bidirectional translocation of membrane-inserted phospholipid probes and to decrease the asymmetric distribution of endogenous membrane phospholipids. These scrambling effects are now shown to be suppressed by pretreatment of cells with the essentially impermeable reagents 4,4'-diisothiocyanostilbene-2,2'-disulfonic ... More