Interaction Analysis of a Two-Component System Using Nanodiscs

DC FieldValueLanguage
dc.contributor.authorHoernschemeyer, Patrick
dc.contributor.authorLiss, Viktoria
dc.contributor.authorHeermann, Ralf
dc.contributor.authorJung, Kirsten
dc.contributor.authorHunke, Sabine
dc.date.accessioned2021-12-23T16:11:00Z-
dc.date.available2021-12-23T16:11:00Z-
dc.date.issued2016
dc.identifier.issn19326203
dc.identifier.urihttps://osnascholar.ub.uni-osnabrueck.de/handle/unios/9489-
dc.description.abstractTwo-component systems are the major means by which bacteria couple adaptation to environmental changes. All utilize a phosphorylation cascade from a histidine kinase to a response regulator, and some also employ an accessory protein. The system-wide signaling fidelity of two-component systems is based on preferential binding between the signaling proteins. However, information on the interaction kinetics between membrane embedded histidine kinase and its partner proteins is lacking. Here, we report the first analysis of the interactions between the full-length membrane-bound histidine kinase CpxA, which was reconstituted in nanodiscs, and its cognate response regulator CpxR and accessory protein CpxP. Using surface plasmon resonance spectroscopy in combination with interaction map analysis, the affinity of membrane-embedded CpxA for CpxR was quantified, and found to increase by tenfold in the presence of ATP, suggesting that a considerable portion of phosphorylated CpxR might be stably associated with CpxA in vivo. Using microscale thermophoresis, the affinity between CpxA in nanodiscs and CpxP was determined to be substantially lower than that between CpxA and CpxR. Taken together, the quantitative interaction data extend our understanding of the signal transduction mechanism used by two-component systems.
dc.description.sponsorshipDFGGerman Research Foundation (DFG)European Commission [Hu1011/2-1, SFB944]; [Exc114/2]; This work was supported by DFG grants Hu1011/2-1 and SFB944 to S.H. as well as Exc114/2 to K.J.; We thank K.-H. Altendorf and Daniel Kummel for continuous support, Michael Hensel, Jacob Piehler and Karl Andersson for fruitful discussions, and E. Limpinsel for technical support. SPR spectroscopy was performed in the Bioanalytics Core Facility at the Ludwig-Maximilians-Universitat Munchen Biocenter. This work was supported by DFG grants Hu1011/2-1 and SFB944 to S.H. as well as Exc114/2 to K.J.
dc.language.isoen
dc.publisherPUBLIC LIBRARY SCIENCE
dc.relation.ispartofPLOS ONE
dc.subjectENVELOPE STRESS-RESPONSE
dc.subjectESCHERICHIA-COLI
dc.subjectHISTIDINE KINASE
dc.subjectINTEGRATION
dc.subjectMICROSCALE THERMOPHORESIS
dc.subjectMultidisciplinary Sciences
dc.subjectPHOSPHORYLATION
dc.subjectPROTEIN
dc.subjectREGULATOR
dc.subjectScience & Technology - Other Topics
dc.subjectSIGNAL-TRANSDUCTION
dc.subjectSPECIFICITY
dc.titleInteraction Analysis of a Two-Component System Using Nanodiscs
dc.typejournal article
dc.identifier.doi10.1371/journal.pone.0149187
dc.identifier.isiISI:000371219000054
dc.description.volume11
dc.description.issue2
dc.contributor.orcid0000-0003-0631-6156
dc.contributor.orcid0000-0002-1304-2512
dc.contributor.researcheridE-1793-2013
dc.publisher.place1160 BATTERY STREET, STE 100, SAN FRANCISCO, CA 94111 USA
dcterms.isPartOf.abbreviationPLoS One
dcterms.oaStatusGreen Submitted, Green Published, gold
crisitem.author.deptFB 05 - Biologie/Chemie-
crisitem.author.deptidfb05-
crisitem.author.parentorgUniversität Osnabrück-
crisitem.author.netidHuSa260-
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