Skip to main content
SpringerLink
Log in

Anion exchange through band 3 protein in canine leishmaniasis at different stages of disease

  • ION CHANNELS, RECEPTORS AND TRANSPORTERS
  • Published:
Pflügers Archiv - European Journal of Physiology Aims and scope Submit manuscript

Abstract

Band 3 protein efficiency in mediating Cl−/HCO3 − exchange through erythrocytes membrane is reduced by oxidative stress. The aim of the present study was to verify whether and how anion transport through band 3 protein may be useful in monitoring canine leishmaniasis (Leishmania infantum) development, a disease associated to membrane protein degradation and oxidative stress. To accomplish this aim, serological analysis to determine IFAT (immunofluorescence antibody test) titers against leishmaniasis has been performed and 1:160 and 1:540 titers, determined at diagnosis and after 6 months, were considered for experiments. Oxidative conditions have been assessed by estimating MDA (malondialdehyde) plasma levels, intracellular GSH (reduced glutathione) content, and membrane –SH groups. Band 3 protein anion exchange capability was evaluated by measuring the rate constant for SO4 = uptake, and its expression levels, along with those of P-Tyr (phosphorylated tyrosine), involved in pathways underlying band 3 protein function, have been also determined. Our results show that, in infected dogs with 1:160 IFAT titer, high MDA plasma levels and oxidation of –SH groups are associated to increased P-Tyr expression levels, leading to a reduction in anion exchange capability throughout 6 months of diagnosis. On the other hand, infected dogs with 1:540 IFAT titer, exhibited oxidative conditions associated to an impaired anion exchange capability at diagnosis, were ameliorated after 6 months. Such findings suggest that (1) band 3 protein-mediated anion transport is reduced by oxidative conditions associated to leishmaniasis, putatively via phosphorylative pathways; (2) band 3 protein efficiency may account for canine leishmaniasis development; and (3) the assessment of band 3 protein function may represent an additional tool for canine leishmaniasis diagnosis and monitoring of its development, with potential application to humans, either in case of leishmaniasis or other oxidative-related pathologies.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Alenghat FJ, Golan DE (2013) Membrane protein dynamics and functional implications in mammalian cells. Curr Top Membr 72:89–120

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Almeida BF, Narciso LG, Melo LM, Preve PP, Bosco AM, Lima VM, Ciarlini PC (2013) Leishmaniasis causes oxidative stress and alteration of oxidative metabolism and viability of neutrophils in dogs. Vet J 198(3):599–605

    Article  CAS  PubMed  Google Scholar 

  3. Arakawa T, Kobayashi-Yurugi T, Alguel Y, Iwanari H, Hatae H, Iwata M, Abe Y, Hino T, Ikeda-Suno C, Kuma H, Kang D, Murata T, Hamakubo T, Cameron AD, Kobayashi T, Hamasaki N, Iwata S (2015) Crystal structure of the anion exchanger domain of human erythrocyte band 3. Science 350:680–684

    Article  CAS  PubMed  Google Scholar 

  4. Bettini S, Gradoni L (1986) Canine Leishmaniasis in the Mediterranean area and in its implications for human leishmaniases. Insect Sci Appl 7:241–245

    Google Scholar 

  5. Biswas T, Pal JK, Naskar K, Ghosh DK, Ghosal J (1995) Lipid peroxidation of erythrocytes during anemia of the hamsters infected with Leishmania donovani. Mol Cell Biochem 146:99–105

    Article  CAS  PubMed  Google Scholar 

  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  7. Brianti E, Gaglio G, Napoli E, Falsone L, Prudente C, Solari Basano F, Latrofa MS, Tarallo VD, Dantas-Torres F, Capelli G, Stanneck D, Giannetto S, Otranto D (2014) Efficacy of a slow-release imidacloprid (10%)/flumethrin (4.5%) collar for the prevention of canine leishmaniosis. Parasit Vectors 7:327

    Article  PubMed  PubMed Central  Google Scholar 

  8. Celedón G, Gonzalez G, Ferrer V, Lissi EA (2004) Peroxyloxidized erythrocyte membrane band 3 protein with anion transport capacity is degraded by membrane-bound proteinase. Free Rad Res 38:1055–1059

    Article  Google Scholar 

  9. Chowdhury KD, Sen G, Biswas T (2010) Regulatory role of nitric oxide in the reduced survival of erythrocytes in visceral leishmaniasis. Biochim Biophys Acta1800(9):964–976

  10. Ciaramella P, Oliva G, Luna RD, Gradoni L, Ambrosio R, Cortese L, Scalone A, Persechino A (1997) A retrospective clinical study of canine leishmaniasis in 150 dogs naturally infected by Leishmania infantum. Vet Rec 141(21):539–543

    Article  CAS  PubMed  Google Scholar 

  11. Çimen MYB (2008) Free radical metabolism in human erythrocytes. Clin Chim Acta 390(1–2):1–11

    Article  PubMed  Google Scholar 

  12. Coura-Vital W, Marques MJ, Giunchetti RC, Teixeira-Carvalho A, Moreira ND, Vitoriano-Souza J, Vieira PM, Carneiro CM, Corrêa-Oliveira R, Martins-Filho OA, Carneiro M, Reis AB (2011) Humoral and cellular immune responses in dogs with inapparent natural Leishmania infantum infection. Vet J 190:43–47

    Article  Google Scholar 

  13. Crupi M, Romano L, Romano P, Venza M, Venza I, Teti D (2010) Erythrocytes anion transport and oxidative change in b-thalassaemias. Cell Biol Int 34:655–662

    Article  Google Scholar 

  14. Dantas-Torres F, Tarallo VD, Latrofa MS, Falchi A, Lia RP, Otranto D (2014) Ecology of phlebotomine sand flies and Leishmania infantum infection in a rural area of southern Italy. Acta Trop 137:67–73

    Article  PubMed  Google Scholar 

  15. De Luna R, Ferrante M, Severino L, Ambrosio R, Piantedosi D, Gradoni L, Lucisano A, Persechino A (2000) Decreased lipid fluidity of the erythrocyte membrane in dogs with leishmaniasis-associated anaemia. J Comp Pathol 122(2–3):213–216

    Article  CAS  PubMed  Google Scholar 

  16. De Tommasi AS, Otranto D, Furlanello T, Tasca S, Cantacessi C, Breitschwerdt EB, Stanneck D, Dantas-Torres F, Baneth G, Capelli G, de Caprariis D (2014) Evaluation of blood and bone marrow in selected canine vector-borne diseases. Parasit Vectors 7:534

    Article  PubMed  PubMed Central  Google Scholar 

  17. Dekowski SA, Rybicki A, Drickamer K (1983) A tyrosine kinase associated with the red cell membrane phosphorylates band 3. J Biol Chem 258:2750–2753

    CAS  PubMed  Google Scholar 

  18. Foglia Manzillo V, Di Muccio T, Cappiello S, Scalone A, Paparcone R, Fiorentino E, Gizzarelli M, Gramiccia M, Gradoni L, Oliva G (2013) Prospective study on the incidence and progression of clinical signs in naïve dogs naturally infected by Leishmania infantum. PLoS Negl Trop Dis 7:e2225

    Article  PubMed  PubMed Central  Google Scholar 

  19. Giustarini D, Dalle-Donne I, Milzani A, Fanti P, Rossi R (2013) Analysis of GSH and GSSG after derivatization with N-ethylmaleimide. Nat Protoc 8(9):1660–1669

    Article  CAS  PubMed  Google Scholar 

  20. Gugliotta T, De Luca G, Romano P, Rigano C, Scuteri A, Romano L (2012) Effects of lead chloride on human erythrocyte membranes and on kinetic anion sulphate and glutathione concentrations. Cell Mol Biol Lett 17(4):586–589

    Article  CAS  PubMed  Google Scholar 

  21. Harrison ML, Isaacson CC, Burg DL, Geahlen RL, Low PS (1994) Phosphorylation of human erythrocyte band 3 by endogenous p72syk. J Biol Chem 269(2):955–959

    CAS  PubMed  Google Scholar 

  22. Jennings ML (1976) Proton fluxes associated with erythrocyte membrane anion exchange. J Membrane Biol 28:187–205

    Article  CAS  Google Scholar 

  23. Jessen F, Sjoholm C, Hoffmann EK (1986) Identification of the anion exchange protein of Ehrlich cells: a kinetic analysis of the inhibitory effects of 4,4′-diisothiocyano-2,2′-stilbene-disulfonic acid (DIDS) and labeling of membrane proteins with 3H-DIDS. J Membrane Biol 92:195–205

    Article  CAS  Google Scholar 

  24. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227(5259):680–685

    Article  CAS  PubMed  Google Scholar 

  25. Low PS, Allen DP, Zioncheck TF, Chari P, Willardson BM, Geahlen RL, Harrison ML (1987) Tyrosine phosphorylation of band 3 inhibits peripheral protein binding. J Biol Chem 262(10):4592–4596

    CAS  PubMed  Google Scholar 

  26. Low FM, Hampton MB, Peskin AV, Winterbourn CC (2007) Peroxiredoxin 2 functions as a noncatalytic scavenger of low-level hydrogen peroxide in the erythrocyte. Blood 109:2611–2617

    Article  CAS  PubMed  Google Scholar 

  27. Macchi B, Marino-Merlo F, Frezza C, Cuzzocrea S, Mastino A (2014) Inflammation and programmed cell death in Alzheimer’s disease: comparison of the central nervous system and peripheral blood. Mol Neurobiol 50(2):463–472

    Article  CAS  PubMed  Google Scholar 

  28. Macchi B, Di Paola R, Marino-Merlo F, Felice MR, Cuzzocrea S, Mastino A (2015) Inflammatory and cell death pathways in brain and peripheral blood in Parkinson’s disease. CNS Neurol Disord Drug Targets 14(3):313–324

    Article  CAS  PubMed  Google Scholar 

  29. Mallozzi C, Di Stasi A, Minetti M (1997) Peroxynitrite modulates tyrosine-dependent signal transduction pathway of human erythrocyte band 3. FASEB J 11:1281–1290

    CAS  PubMed  Google Scholar 

  30. Morabito R, Marino A, Romano P, Rigano C, La Spada G (2013) Sulphate and chloride-dependent potassium transport in human erythrocytes are affected by crude venom from nematocysts of the jellyfish Pelagia noctiluca. Cell Physiol Biochem 32:86–95

    Article  CAS  PubMed  Google Scholar 

  31. Morabito R, Falliti G, Geraci A, La Spada G, Marino A (2015) Curcumin protects –SH groups and sulphate transport after oxidative damage in human erythrocytes. Cell Physiol Biochem 36:345–357

    Article  CAS  PubMed  Google Scholar 

  32. Morabito R, La Spada G, Crupi R, Esposito E, Marino A (2015) Crude venom from nematocysts of the jellyfish Pelagia noctiluca as a tool to study cell physiology. CNSAMC (Central Nervous System Agents in Medicinal Chemistry) special issue ‘The activity of products from Cnidaria: a therapeutic tool in neurological diseases?’ 15(2):68–73

  33. Morabito R, Romano O, La Spada G, Marino A (2016) H2O2-induced oxidative stress affects SO4 = transport in human erythrocytes. PLoS One 11(1):e0146485

    Article  PubMed  PubMed Central  Google Scholar 

  34. Morabito R, Remigante A, Di Pietro ML, Giannetto A, La Spada G, Marino A (2017) SO4 = uptake and catalase role in preconditioning after H2O2-induced oxidative stress in human erythrocytes. Pflügers Arch 469(2):235–250

    Article  CAS  PubMed  Google Scholar 

  35. Naoum PC, Radispiel J, Moraes M da Silva (2004) Spectrometric measurement of methemoglobin without interference of chemical or enzymatic reagents. Rev Bras Hematol Hemoter 26:19–22

  36. Nejjar R, Lemrani M, Boucedda L, Amarouch H, Benslimane A (2000) Variation in antibody titres against Leishmania infantum in naturally infected dogs in northern Morocco. Rev Med Vet 151:841–846

    Google Scholar 

  37. Nicolato Rde C, de Abreu RT, Roatt BM, Aguiar-Soares RD, Reis LE, Md C, Carneiro CM, Giunchetti RC, Bouillet LE, Lemos DS, Coura-Vital W, Reis AB (2013) Clinical forms of canine visceral Leishmaniasis in naturally Leishmania infantum-infected dogs and related myelogram and hemogram changes. PLoS One 8(12):e82947

    Article  PubMed  Google Scholar 

  38. Nieto CG, García-Alonso M, Requena JM, Mirón C, Soto M, Alonso C, Navarrete I (1999) Analysis of the humoral immune response against total and recombinant antigens of Leishmania infantum: correlation with disease progression in canine experimental leishmaniasis. Vet Immunol Immunopathol 67(2):117–130

    Article  CAS  PubMed  Google Scholar 

  39. Pantaleo A, Giribaldi G, Mannu F, Arese P, Turrini F (2008) Naturally occurring anti-band 3 antibodies and red blood cell removal under physiological and pathological conditions. Autoimmun Rev 7:457–462

    Article  CAS  PubMed  Google Scholar 

  40. Pantaleo A, Ferru E, Pau MC, Khadjavi A, Mandili G, Mattè A, Spano A, De Franceschi L, Pippia P, Turrini F (2016) Band 3 erythrocyte membrane protein acts as redox stress sensor leading to its phosphorylation by p (72) Syk. Oxidative Med Cell Longev 2016:6051093

    Article  Google Scholar 

  41. Passow H (1986) Molecular aspects of band 3 protein-mediated anion transport across the red blood cell membrane. Rev Physiol Biochem Pharmacol 103:61–203

    CAS  PubMed  Google Scholar 

  42. Pinelli E, Killick-Kendrick R, Wagenaar J, Bernadina W, Del Real G, Ruitenberg J (1994) Cellular and humoral immune responses in dogs experimentally and naturally infected with Leishmania infantum. Infect Immun 62:229–235

    CAS  PubMed  PubMed Central  Google Scholar 

  43. Proverbio D, Spada E, Bagnagatti de Giorgi G, Perego R, Valena E (2014) Relationship between Leishmania IFAT titer and clinicopathological manifestations (clinical score) in dogs. Biomed Res Int 412808:1–5

    Article  Google Scholar 

  44. Reddy YN, Murthy SV, Krishna DR, Prabhakar MC (2004) Role of free radicals and antioxidants in tuberculosis patients. Indian J Tuberc 51:213–218

    Google Scholar 

  45. Reithmeier RAF, Casey JR, Kall AC, Sansomc MSP, Alguel Y, Iwata S (2016) Band 3, the human red cell chloride/bicarbonate anion exchanger (AE1, SLC4A1), in a structural context. Biochim Biophys Acta 1858:1507–1532

    Article  CAS  PubMed  Google Scholar 

  46. Reithmer RAF (1993) The erythrocytes anion transporter (band 3). Curr Opin Struct Biol 3:515–523

    Article  Google Scholar 

  47. Romano L, Passow H (1984) Characterization of anion transport system in trout red blood cell. Am J Phys 246:C330–C338

    CAS  Google Scholar 

  48. Romano L, Scuteri A, Gugliotta T, Romano P, De Luca G, Sidoti A, Amato A (2002) Sulphate influx in the erythrocytes of normotensive, diabetic and hypertensive patients. Cell Biol Int 26(5):421–426

    Article  CAS  PubMed  Google Scholar 

  49. Roy SS, Sen G, Biswas T (2005) Role of sulfhydryl groups in band 3 in the inhibition of phosphate transport across erythrocyte membrane in visceral leishmaniasis. Arch Biochem Biophys 436:121–127

    Article  CAS  PubMed  Google Scholar 

  50. Sen G, Ghosal J, Naskar K, Biswas T (2000) Altered calcium homeostasis and membrane destabilization in erythrocytes of hamsters infected with Leishmania donovani. Ann Trop Med Parasitol 93:43–53

    Article  Google Scholar 

  51. Sen G, Mukhopadhyay R, Ghosal J, Biswas T (2001) Oxidative damage of erythrocytes: a possible mechanism for premature hemolysis in experimental visceral leishmaniasis in hamsters. Ann Hematol 80:32–37

    Article  CAS  PubMed  Google Scholar 

  52. Sen G, Mandal S, Roy SS, Mukhopadhyay S, Biswas T (2005) Therapeutic use of quercetin in the control of infection and anemia associated with visceral leishmaniasis. Free Radic Biol Med 38:1257–1264

    Article  CAS  PubMed  Google Scholar 

  53. Steck TL (1974) The organization of proteins in the human red blood cell membrane. A review. J Cell Biol 62:1–19

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Teti D, Crupi M, Busà M, Valenti A, Loddo S, Mondello M, Romano L (2005) Chemical and pathological oxidative influences on band 3 protein anion-exchanger. Cell Physiol Biochem 16:77–86

    Article  CAS  PubMed  Google Scholar 

  55. Tropia de Abreu R, Carvalho MG, Carneiro CM, Giunchetti RC, Teixeira-Carvalho A, Martins-Filho OA, Coura-Vital W, Corrêa-Oliveira R, Reis AB (2011) Influence of clinical status and parasite load on erythropoiesis and leucopoiesis in dogs naturally infected with Leishmania (Leishmania) chagasi. PLoS One 6:e18873

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Vercammen F, Fernandez-Perez FJ, Del Amo C, Alunda JM (2002) Follow-up of Leishmania infantum naturally infected dogs treated with allopurinol: immunofluorescence antibody test, ELISA and Western blot. Acta Trop 84(3):175–181

    Article  CAS  PubMed  Google Scholar 

  57. Veskoukis AS, Kyparos A, Paschalis V, Nikolaidis MG (2016) Spectrophotometric assays for measuring redox biomarkers in blood. Biomarkers 21(3):208–217

    Article  CAS  PubMed  Google Scholar 

  58. Welbourn EM, Wilson MT, Yusof A, Metodiev MV, Cooper CE (2017) The mechanism of formation, structure and physiological relevance of covalent hemoglobin attachment to the erythrocyte membrane. Free Radic Biol Med 103:95–106

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Xiang W, Weisbach V, Sticht H, Seebahn A, Bussmann J, Zimmermann R, Becker CM (2013) Oxidative stress-induced posttranslational modifications of human hemoglobin in erythrocytes. Arch Biochem Biophys 529(1):34–44

    Article  CAS  PubMed  Google Scholar 

  60. Yannoukakos D, Vasseur C, Piau JP, Wajeman H, Bursaux E (1991) Phosphorylation sites in human erythrocyte band 3 protein. Biochim Biophys Acta 1061:253–266

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors are grateful to Dr. Fabrizio Vitale from the Centro di referenza nazionale Leishmaniosi, Istituto Zooprofilattico della Sicilia “A. Mirri,” Palermo (Italy), for serological analysis and Prof. L. Romano for encouragement and manuscript revision.

Author information

Author notes

  1. Rossana Morabito and Alessia Remigante contributed equally to this work.

Authors and Affiliations

  1. Department of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, Viale F. Stagno D’Alcontres 31, 98166, Messina, Italy

    Rossana Morabito, Alessia Remigante, Giuseppina La Spada & Angela Marino

  2. Department of Veterinary Sciences, Polo Universitario dell’Annunziata, 98168, Messina, Italy

    Mauro Cavallaro

  3. Veterinary, Messina, Italy

    Alessandro Taormina

Authors
  1. Rossana Morabito
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alessia Remigante
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mauro Cavallaro
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Alessandro Taormina
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Giuseppina La Spada
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Angela Marino
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Angela Marino.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Electronic supplementary material

ESM 1

(PPTX 189 kb).

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Morabito, R., Remigante, A., Cavallaro, M. et al. Anion exchange through band 3 protein in canine leishmaniasis at different stages of disease. Pflugers Arch - Eur J Physiol 469, 713–724 (2017). https://doi.org/10.1007/s00424-017-1974-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00424-017-1974-2

Keywords

Search

Navigation