Dynamic interactions of CbiN and CbiM trigger activity of a cobalt energy-coupling-factor transporter
DC Element | Wert | Sprache |
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dc.contributor.author | Finkenwirth, Friedrich | |
dc.contributor.author | Sippach, Michael | |
dc.contributor.author | Pecina, Sinah N. | |
dc.contributor.author | Gaede, Mario | |
dc.contributor.author | Ruta, Julia | |
dc.contributor.author | Ricke, Adrian | |
dc.contributor.author | Bondarenko, Elena | |
dc.contributor.author | Klare, Johann P. | |
dc.contributor.author | Zinke, Maximilian | |
dc.contributor.author | Lange, Sascha | |
dc.contributor.author | Lange, Adam | |
dc.contributor.author | Steinhoff, Heinz-Jurgen | |
dc.contributor.author | Eitinger, Thomas | |
dc.date.accessioned | 2021-12-23T16:12:14Z | - |
dc.date.available | 2021-12-23T16:12:14Z | - |
dc.date.issued | 2020 | |
dc.identifier.issn | 00052736 | |
dc.identifier.uri | https://osnascholar.ub.uni-osnabrueck.de/handle/unios/10113 | - |
dc.description.abstract | Energy-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.sponsorship | Deutsche 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.iso | en | |
dc.publisher | ELSEVIER | |
dc.relation.ispartof | BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES | |
dc.subject | ATP-binding cassette transporter | |
dc.subject | Biochemistry & Molecular Biology | |
dc.subject | Biophysics | |
dc.subject | Electron paramagnetic resonance (EPR) | |
dc.subject | EPR | |
dc.subject | MECHANISM | |
dc.subject | Metal ion-protein interaction | |
dc.subject | MOTION | |
dc.subject | NICKEL | |
dc.subject | PERMEASES | |
dc.subject | PREDICTION | |
dc.subject | Protein conformation | |
dc.subject | Protein-protein interaction | |
dc.subject | PROTEINS | |
dc.subject | Solid-state NMR | |
dc.subject | SPECIFICITY | |
dc.subject | SPIN | |
dc.subject | SUBSTRATE CAPTURE | |
dc.title | Dynamic interactions of CbiN and CbiM trigger activity of a cobalt energy-coupling-factor transporter | |
dc.type | journal article | |
dc.identifier.doi | 10.1016/j.bbamem.2019.183114 | |
dc.identifier.isi | ISI:000509632200046 | |
dc.description.volume | 1862 | |
dc.description.issue | 2 | |
dc.contributor.orcid | 0000-0002-5761-5968 | |
dc.contributor.orcid | 0000-0002-5888-0157 | |
dc.contributor.orcid | 0000-0002-0070-8725 | |
dc.contributor.orcid | 0000-0002-7534-5973 | |
dc.contributor.orcid | 0000-0002-0541-5139 | |
dc.contributor.researcherid | C-1428-2009 | |
dc.contributor.researcherid | H-3791-2014 | |
dc.identifier.eissn | 18792642 | |
dc.publisher.place | RADARWEG 29, 1043 NX AMSTERDAM, NETHERLANDS | |
dcterms.isPartOf.abbreviation | Biochim. Biophys. Acta-Biomembr. | |
dcterms.oaStatus | hybrid | |
crisitem.author.dept | FB 04 - Physik | - |
crisitem.author.deptid | fb04 | - |
crisitem.author.parentorg | Universität Osnabrück | - |
crisitem.author.netid | StHe633 | - |
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geprüft am 01.06.2024