Association Properties and Unfolding of a βγ-Crystallin Domain of a Vibrio-Specific Protein.docVIP

AssociationPropertiesandUnfoldingofabc-Crystallin DomainofaVibrio-SpecificProtein ShashiKumarSuman,DaddaliRavindra,YogendraSharma*,AmitaMishra* CentreforCellularandMolecularBiology,CouncilofScientificandIndustrialResearch,UppalRoad,Hyderabad,India Abstract Thebc-crystallinsuperfamilypossessesalargenumberofversatilemembers,ofwhichonlyafewmembersotherthanlens bc-crystallinshavebeenstudied.Understandingthenon-crystallinfunctionsaswellasoriginofcrystallin-likepropertiesof suchproteinsispossiblebyexploringnovelmembersfromdiversesources.Wedescribeanovelbc-crystallindomainwith S-type(Spherulin3atype)GreekkeymotifsinproteinvibrillinfromapathogenicbacteriumVibriocholerae.Thisdomainisa partofalargeVibrio-specificproteinprevalentinVibriospecies(foundinatleastfourteendifferentstrainssequencedso 2+ 2+ far). The domain contains two canonical N/D-N/D-X-X-S/T-S Ca -binding motifs, and bind Ca . Unlike spherulin 3a and othermicrobialhomologuesstudiedsofar,bc-crystallindomainofvibrillinself-associatesformingoligomersofvarioussizes including dimers. The fractionated dimers readily form octamers in concentration-dependent manner, suggesting an associationbetweenthesetwomajorforms.Thedomainassociates/dissociatesformingdimersatthecostofmonomeric 2+ 2+ populations in the presence of Ca . No such effect of Ca has been observed in oligomeric species. The equilibrium unfolding of both forms follows a similar pattern, with the formation of an unfolding intermediate at sub-molar concentrationsofdenaturant.Thesepropertiesexhibitedbythisbc-crystallindomainarenotshownbyanyotherdomain studiedsofar,demonstratingthediversityindomainproperties. Citation: SumanSK,Ravindra D,SharmaY,MishraA(2013)AssociationPropertiesandUnfoldingofabc-CrystallinDomainofaVibrio-Specific Protein.PLoS ONE8(1):e53610.doi:10.1371/journal.pone.0053610 Editor:YangZhang,UniversityofMichigan,UnitedStatesofAmerica ReceivedJuly19,2011;AcceptedDecember3,2012;PublishedJanuary22,2013 Copyright: ? 2013 Suman et al. This is