Structural and biophysical analysis of a Haemophilus influenzae tripartite ATP-independent periplasmic (TRAP) transporter

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dc.contributor.authorCurrie, Michael J.
dc.contributor.authorDavies, James S.
dc.contributor.authorScalise, Mariafrancesca
dc.contributor.authorGulati, Ashutosh
dc.contributor.authorWright, Joshua D.
dc.contributor.authorNewton-Vesty, Michael C.
dc.contributor.authorAbeysekera, Gayan S.
dc.contributor.authorSubramanian, Ramaswamy
dc.contributor.authorWahlgren, Weixiao Y.
dc.contributor.authorFriemann, Rosmarie
dc.contributor.authorAllison, Jane R.
dc.contributor.authorMace, Peter D.
dc.contributor.authorGriffin, Michael D. W.
dc.contributor.authorDemeler, Borries
dc.contributor.authorWakatsuki, Soichi
dc.contributor.authorDrew, David
dc.contributor.authorIndiveri, Cesare
dc.contributor.authorDobson, Renwick C. J.
dc.contributor.authorNorth, Rachel A.
dc.date.accessioned2024-07-11T20:11:19Z
dc.date.available2024-07-11T20:11:19Z
dc.date.issued2024
dc.descriptionOpen access article. Creative Commons Attribution 4.0 International license (CC BY 4.0) applies
dc.description.abstractTripartite ATP-independent periplasmic (TRAP) transporters are secondary-active transporters that receive their substrates via a soluble-binding protein to move bioorganic acids across bacterial or archaeal cell membranes. Recent cryo-electron microscopy (cryo-EM) structures of TRAP transporters provide a broad framework to understand how they work, but the mechanistic details of transport are not yet defined. Here we report the cryo-EM structure of the Haemophilus influenzae N-acetylneuraminate TRAP transporter (HiSiaQM) at 2.99 Å resolution (extending to 2.2 Å at the core), revealing new features. The improved resolution (the previous HiSiaQM structure is 4.7 Å resolution) permits accurate assignment of two Na+ sites and the architecture of the substrate-binding site, consistent with mutagenic and functional data. Moreover, rather than a monomer, the HiSiaQM structure is a homodimer. We observe lipids at the dimer interface, as well as a lipid trapped within the fusion that links the SiaQ and SiaM subunits. We show that the affinity (KD) for the complex between the soluble HiSiaP protein and HiSiaQM is in the micromolar range and that a related SiaP can bind HiSiaQM. This work provides key data that enhances our understanding of the ‘elevator-with-an-operator’ mechanism of TRAP transporters.
dc.description.peer-reviewYes
dc.identifier.citationCurrie, M. J., Davies, J. S., Scalise, M., Gulati, A., Wright, J. D., Newton-Vesty, M. C., Abeysekera, G. S., Subramanian, R., Wahlgren, W. Y., Friemann, R., Allison, J. R., Mace, P. D., Griffin, M. D. W., Demeler, B., Wakatsuki, S., Drew, D., Indiveri, C., Dobson, R. C. J., & North, R. A. (2024). Structural and biophysical analysis of a Haemophilus influenzae tripartite ATP-independent periplasmic (TRAP) transporter. eLife, 12, Article RP92307. https://doi.org/10.7554/eLife.92307.3
dc.identifier.urihttps://hdl.handle.net/10133/6816
dc.language.isoen
dc.publishereLife Sciences Publications
dc.publisher.departmentDepartment of Chemistry and Biochemistry
dc.publisher.facultyArts and Science
dc.publisher.institutionUniversity of Canterbury
dc.publisher.institutionStockholm University
dc.publisher.institutionUniversity of Calabria
dc.publisher.institutionPurdue University West Lafayette
dc.publisher.institutionUniversity of Gothenburg
dc.publisher.institutionUniversity of Auckland
dc.publisher.institutionUniversity of Otago
dc.publisher.institutionUniversity of Melbourne
dc.publisher.institutionUniversity of Montana
dc.publisher.institutionUniversity of Lethbridge
dc.publisher.institutionSLAC National Accelerator Laboratory
dc.publisher.institutionStanford University School of Medicine
dc.publisher.institutionCNR Institute of Biomembranes
dc.publisher.institutionUniversity of Sydney
dc.publisher.urlhttps://doi.org/10.7554/eLife.92307.3
dc.subjectNeu5Ac
dc.subjectMembrane transport proteins
dc.subjectMolecular biophysics
dc.subjectProtein-protein interaction
dc.subjectSialic acid
dc.subjectStructural biology
dc.subjectTransport mechanism
dc.subject.lcshMolecular biology
dc.subject.lcshProtein-protein interactions
dc.subject.lcshSialic acids
dc.subject.lcshCarrier proteins
dc.titleStructural and biophysical analysis of a Haemophilus influenzae tripartite ATP-independent periplasmic (TRAP) transporter
dc.typeArticle
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