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dc.contributor.authorKojetin, D. J.
dc.contributor.authorThompson, R. J.
dc.contributor.authorBenson, L. M.
dc.contributor.authorNaylor, S.
dc.contributor.authorWaterman, J.
dc.contributor.authorDavies, Keith
dc.contributor.authorOpperman, Charles H.
dc.contributor.authorStephenson, Keith
dc.contributor.authorHoch, J. A.
dc.contributor.authorCavanagh, J.
dc.date.accessioned2013-06-05T12:15:51Z
dc.date.available2013-06-05T12:15:51Z
dc.date.issued2005-10
dc.identifier.citationKojetin , D J , Thompson , R J , Benson , L M , Naylor , S , Waterman , J , Davies , K , Opperman , C H , Stephenson , K , Hoch , J A & Cavanagh , J 2005 , ' Structural analysis of divalent metals binding to the Bacillus subtilis response regulator Spo0F : the possibility for In vitro metalloregulation in the initiation of sporulation ' , Biometals , vol. 18 , no. 5 , pp. 449-466 . https://doi.org/10.1007/s10534-005-4303-8
dc.identifier.issn0966-0844
dc.identifier.otherPURE: 590000
dc.identifier.otherPURE UUID: 050ff9cb-f55f-4a50-b2cd-e32dd5bcc06e
dc.identifier.otherWOS: 000233823500001
dc.identifier.otherScopus: 28644440751
dc.identifier.otherORCID: /0000-0001-6060-2394/work/32215798
dc.identifier.urihttp://hdl.handle.net/2299/10717
dc.description.abstractThe presence of a divalent metal ion in a negatively charged aspartic acid pocket is essential for phosphorylation of response regulator proteins. Here, we present metal binding studies of the Bacillus subtilis response regulator Spo0F using NMR and mu ESI-MS. NMR studies show that the divalent metals Ca2+, Mg2+ and Mn2+ primarily bind, as expected, in the Asp pocket phosphorylation site. However, identical studies with Cu2+ show distinct binding effects in three specific locations: (i) the Asp pocket, (ii) a grouping of charged residues at a site opposite of the Asp pocket, and (iii) on the beta 4-alpha 4 loop and the beta 5/alpha 5 interface, particularly around and including H101. mu ESI-MS studies stoichiometrically confirm the NMR studies and demonstrate that most divalent metal ions bind to Spo0F primarily in a 1:1 ratio. Again, in the case of Cu2+, multiple metal-bound species are observed. Subsequent experiments reveal that Mg2+ supports phosphotransfer between KinA and Spo0F, while Cu2+ fails to support KinA phosphotransfer. Additionally, the presence of Cu2+ at non-lethal concentrations in sporulation media for B. subtilis and the related organism Pasteuria penetrans was found to inhibit spore formation while continuing to permit vegetative growth. Depending on the type of divalent metal ion present, in vitro phosphorylation of Spo0F by its cognate kinase KinA can be inhibited.en
dc.format.extent18
dc.language.isoeng
dc.relation.ispartofBiometals
dc.subjectmetal binding
dc.subjectNMR spectroscopy
dc.subjectresponse regulator
dc.subjectsporulation
dc.subjecttwo-component signal transduction
dc.subjectHIGH-RESOLUTION NMR
dc.subjectMOLECULAR RECOGNITION
dc.subjectSIGNAL-TRANSDUCTION
dc.subjectPHOSPHORELAY
dc.subjectPROTEINS
dc.subjectSPOOF
dc.subjectPHOSPHOTRANSFER
dc.subjectMEGATERIUM
dc.subjectREVEALS
dc.subjectPHOSPHORYLATION
dc.titleStructural analysis of divalent metals binding to the Bacillus subtilis response regulator Spo0F : the possibility for In vitro metalloregulation in the initiation of sporulationen
dc.contributor.institutionDepartment of Human and Environmental Sciences
dc.contributor.institutionHealth & Human Sciences Research Institute
dc.description.statusPeer reviewed
rioxxterms.versionofrecordhttps://doi.org/10.1007/s10534-005-4303-8
rioxxterms.typeJournal Article/Review
herts.preservation.rarelyaccessedtrue


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