Band 3(B3),theaniontransporter,isanintegralmembraneproteinthatplaysakeystructuralroleby anchoringtheplasmamembranetothespectrin-basedmembraneskeletonintheredcell.Inaddition, it alsoplaysacriticalroleintheassemblyofglycolyticenzymestoregulateredcellmetabolism. However,itsabilitytorecruitproteinsthatcanpreventmembraneoxidationhasnotbeenpreviously explored.Inthisstudy,usingavarietyofexperimentalapproachesincludingcross-linkingstudies, fluorescenceanddichroicmeasurements,surfaceplasmonresonanceanalysis,andproteolyticdigestion assays,wedocumentthattheantioxidantproteinperoxiredoxin-2(PRDX2),thethirdmostabundant cytoplasmicproteininRBCs,interactswiththecytoplasmicdomainofB3.Thesurfaceelectrostatic potentialanalysisandstoichiometrymeasurementsrevealedthattheN-terminalpeptideofB3is involvedintheinteraction.PRDX2underwentaconformationalchangeuponitsbindingtoB3without losingitsperoxidaseactivity.HemichromeformationinducedbyphenylhydrazineofRBCsprevented membraneassociationofPRDX2,implyingoverlappingbindingsites.Documentationoftheabsenceof bindingofPRDX2toB3Neapolisredcellmembranes,inwhichtheinitialN-terminal11aminoacidsare deleted,enabledustoconcludethatPRDX2bindstotheN-terminalcytoplasmicdomainofB3andthat the first11aminoacidsofthisdomainarecrucialforPRDX2membraneassociationinintactRBCs. ThesefindingsimplyyetanotherimportantroleforB3inregulatingredcellmembranefunction.

Membrane association of peroxiredoxin-2 in red cells is mediated by the N-terminal cytoplasmic domain of band3

MATTE', Alessandro;BERTOLDI, Mariarita;SICILIANO, Angela;DE FRANCESCHI, Lucia
2013

Abstract

Band 3(B3),theaniontransporter,isanintegralmembraneproteinthatplaysakeystructuralroleby anchoringtheplasmamembranetothespectrin-basedmembraneskeletonintheredcell.Inaddition, it alsoplaysacriticalroleintheassemblyofglycolyticenzymestoregulateredcellmetabolism. However,itsabilitytorecruitproteinsthatcanpreventmembraneoxidationhasnotbeenpreviously explored.Inthisstudy,usingavarietyofexperimentalapproachesincludingcross-linkingstudies, fluorescenceanddichroicmeasurements,surfaceplasmonresonanceanalysis,andproteolyticdigestion assays,wedocumentthattheantioxidantproteinperoxiredoxin-2(PRDX2),thethirdmostabundant cytoplasmicproteininRBCs,interactswiththecytoplasmicdomainofB3.Thesurfaceelectrostatic potentialanalysisandstoichiometrymeasurementsrevealedthattheN-terminalpeptideofB3is involvedintheinteraction.PRDX2underwentaconformationalchangeuponitsbindingtoB3without losingitsperoxidaseactivity.HemichromeformationinducedbyphenylhydrazineofRBCsprevented membraneassociationofPRDX2,implyingoverlappingbindingsites.Documentationoftheabsenceof bindingofPRDX2toB3Neapolisredcellmembranes,inwhichtheinitialN-terminal11aminoacidsare deleted,enabledustoconcludethatPRDX2bindstotheN-terminalcytoplasmicdomainofB3andthat the first11aminoacidsofthisdomainarecrucialforPRDX2membraneassociationinintactRBCs. ThesefindingsimplyyetanotherimportantroleforB3inregulatingredcellmembranefunction.
peroxiredoxin-2; band 3; red cell membrane; free radicals
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11562/494762
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