Functional Insight into the C-Terminal Extension of Halolysin SptA from Haloarchaeon Natrinema sp. J7 英文参考文献.docVIP

FunctionalInsightintotheC-TerminalExtensionof HalolysinSptAfromHaloarchaeonNatrinemasp.J7 ZhishengXu,XinDu,TingtingLi,FeiGan,BingTang,Xiao-FengTang* StateKeyLaboratoryofVirology,CollegeofLifeSciences,WuhanUniversity,Wuhan,China Abstract HalolysinSptAfromhaloarchaeonNatrinemasp.J7consistsofasubtilisin-likecatalyticdomainandaC-terminalextension (CTE) containing two cysteine residues. In this report, we have investigated the function of the CTE using recombinant enzymesexpressedinHaloferaxvolcaniiWFD11.DeletionoftheCTEgreatlyreducedbutdidnotabolishproteaseactivity, which suggests that the CTE is not essential for enzyme folding. Mutational analysis suggests that residues Cys303 and Cys338 within the CTE form a disulfide bond that make this domain resistant to autocleavage and proteolysis under hypotonicconditions.Characterizationoffull-lengthandCTE-truncationenzymesindicatestheCTEnotonlyconfersextra stability to the enzyme but also assists enzyme activity on protein substrates by facilitating binding at high salinities. Interestingly, homology modeling of the CTE yields a b-jelly roll-like structure similar to those seen in Claudin-binding domain of Clostridium perfringens enterotoxin (clostridial C-CPE) and collagen binding domain (CBD), and the CTE also possessescollagen-bindingactivity,makingitapotentialcandidateasananchoringunitindrugdeliverysystems. Citation:XuZ,DuX,LiT,GanF,TangB,etal.(2011)FunctionalInsightintotheC-TerminalExtensionofHalolysinSptAfromHaloarchaeonNatrinemasp.J7.PLoS ONE6(8):e23562.doi:10.1371/journal.pone.0023562 Editor:VladimirN.Uversky,UniversityofSouthFloridaCollegeofMedicine,UnitedStatesofAmerica ReceivedJune17,2011;AcceptedJuly20,2011;PublishedAugust19,2011 Copyright:?2011Xuetal.Thisisanopen-accessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,whichpermitsunrestricted use,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:ThisworkwassupportedinpartbytheNationalNaturalScienc