Dynamic interactions of CbiN and CbiM trigger activity of a cobalt energy-coupling-factor transporter

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dc.contributor.authorFinkenwirth, Friedrich
dc.contributor.authorSippach, Michael
dc.contributor.authorPecina, Sinah N.
dc.contributor.authorGaede, Mario
dc.contributor.authorRuta, Julia
dc.contributor.authorRicke, Adrian
dc.contributor.authorBondarenko, Elena
dc.contributor.authorKlare, Johann P.
dc.contributor.authorZinke, Maximilian
dc.contributor.authorLange, Sascha
dc.contributor.authorLange, Adam
dc.contributor.authorSteinhoff, Heinz-Jurgen
dc.contributor.authorEitinger, Thomas
dc.date.accessioned2021-12-23T16:12:14Z-
dc.date.available2021-12-23T16:12:14Z-
dc.date.issued2020
dc.identifier.issn00052736
dc.identifier.urihttps://osnascholar.ub.uni-osnabrueck.de/handle/unios/10113-
dc.description.abstractEnergy-coupling factor (ECF) transporters for uptake of vitamins and transition-metal ions into prokaryotic cells share a common architecture consisting of a substrate-specific integral membrane protein (S), a transmembrane coupling protein (T) and two cytoplasmic ATP-binding-cassette-family ATPases. S components rotate within the membrane to expose their binding pockets alternately to the exterior and the cytoplasm. In contrast to vitamin transporters, metal-specific systems rely on additional proteins with essential but poorly understood functions. CbiN, a membrane protein composed of two transmembrane helices tethered by an extracytoplasmic loop of 37 amino-acid residues represents the auxiliary component that temporarily interacts with the CbiMQO(2) Co2+ transporter. CbiN was previously shown to induce significant Co2+ transport activity in the absence of CbiQO(2) in cells producing the S component CbiM plus CbiN or a Cbi(MN) fusion. Here we analyzed the mode of interaction between the two protein domains. Any deletion in the CbiN loop abolished transport activity. In silico predicted protein-protein contacts between segments of the CbiN loop and loops in CbiM were confirmed by cysteine-scanning mutagenesis and crosslinking. Likewise, an ordered structure of the CbiN loop was observed by electron paramagnetic resonance analysis after site-directed spin labeling. The N-terminal loop of CbiM containing three of four metal ligands was partially immobilized in wild-type Cbi(MN) but completely immobile in inactive variants with CbiN loop deletions. Decreased dynamics of the inactive form was also detected by solid-state nuclear magnetic resonance of isotope-labeled protein in proteoliposomes. In conclusion, CbiM-CbiN loop-loop interactions facilitate metal insertion into the binding pocket.
dc.description.sponsorshipDeutsche Forschungsgemeinschaft through PaketantragGerman Research Foundation (DFG) [PAK459, E1374/4-2, STE 640/10, E1374/5-1]; Leibniz-Forschungsinstitut fur Molekulare Pharmakologie (FMP); Funding was provided by the Deutsche Forschungsgemeinschaft through Paketantrag PAK459 by grants E1374/4-2 (to T. E.) and STE 640/10 (to H.-J. S.), and through grant E1374/5-1 (to T. E.). We thank the Leibniz-Forschungsinstitut fur Molekulare Pharmakologie (FMP) for financial support.
dc.language.isoen
dc.publisherELSEVIER
dc.relation.ispartofBIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
dc.subjectATP-binding cassette transporter
dc.subjectBiochemistry & Molecular Biology
dc.subjectBiophysics
dc.subjectElectron paramagnetic resonance (EPR)
dc.subjectEPR
dc.subjectMECHANISM
dc.subjectMetal ion-protein interaction
dc.subjectMOTION
dc.subjectNICKEL
dc.subjectPERMEASES
dc.subjectPREDICTION
dc.subjectProtein conformation
dc.subjectProtein-protein interaction
dc.subjectPROTEINS
dc.subjectSolid-state NMR
dc.subjectSPECIFICITY
dc.subjectSPIN
dc.subjectSUBSTRATE CAPTURE
dc.titleDynamic interactions of CbiN and CbiM trigger activity of a cobalt energy-coupling-factor transporter
dc.typejournal article
dc.identifier.doi10.1016/j.bbamem.2019.183114
dc.identifier.isiISI:000509632200046
dc.description.volume1862
dc.description.issue2
dc.contributor.orcid0000-0002-5761-5968
dc.contributor.orcid0000-0002-5888-0157
dc.contributor.orcid0000-0002-0070-8725
dc.contributor.orcid0000-0002-7534-5973
dc.contributor.orcid0000-0002-0541-5139
dc.contributor.researcheridC-1428-2009
dc.contributor.researcheridH-3791-2014
dc.identifier.eissn18792642
dc.publisher.placeRADARWEG 29, 1043 NX AMSTERDAM, NETHERLANDS
dcterms.isPartOf.abbreviationBiochim. Biophys. Acta-Biomembr.
dcterms.oaStatushybrid
crisitem.author.deptFB 04 - Physik-
crisitem.author.deptidfb04-
crisitem.author.parentorgUniversität Osnabrück-
crisitem.author.netidStHe633-
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