Show simple item record

dc.contributor.authorKumar, Hemant
dc.contributor.authorFiner-Moore, Janet
dc.contributor.authorSmirnova, Irina
dc.contributor.authorKasho, Vladimir
dc.contributor.authorPardon, Els
dc.contributor.authorSteyaert, Jan
dc.contributor.authorKaback, H. Ronald
dc.contributor.authorStroud, Robert M.
dc.date.accessioned2020-05-08T03:13:17Z
dc.date.available2020-05-08T03:13:17Z
dc.date.issued2020-05-07
dc.date.submitted2020-02-06
dc.identifier.otherpone-d-20-03505
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/305143
dc.descriptionFunder: research foundation-flanders
dc.description.abstractThe structure of lactose permease, stabilized in a periplasmic open conformation by two Gly to Trp replacements (LacYww) and complexed with a nanobody directed against this conformation, provides the highest resolution structure of the symporter. The nanobody binds in a different manner than two other nanobodies made against the same mutant, which also bind to the same general region on the periplasmic side. This region of the protein may represent an immune hotspot. The CDR3 loop of the nanobody is held by hydrogen bonds in a conformation that partially blocks access to the substrate-binding site. As a result, kon and koff for galactoside binding to either LacY or the double mutant complexed with the nanobody are lower than for the other two LacY/nanobody complexes though the Kd values are similar, reflecting the fact that the nanobodies rigidify structures along the pathway. While the wild-type LacY/nanobody complex clearly stabilizes a similar ‘extracellular open’ conformation in solution, judged by binding kinetics, the complex with wild-type LacY did not yet crystallize, suggesting the nanobody does not bind strongly enough to shift the equilibrium to stabilize a periplasmic side-open conformation suitable for crystallization. However, the similarity of the galactoside binding kinetics for the nanobody-bound complexes with wild type LacY and with LacYWW indicates that they have similar structures, showing that the reported co-structures reliably show nanobody interactions with LacY.
dc.languageen
dc.publisherPublic Library of Science
dc.rightsAttribution 4.0 International (CC BY 4.0)en
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en
dc.subjectResearch Article
dc.subjectPhysical sciences
dc.subjectBiology and life sciences
dc.titleDiversity in kinetics correlated with structure in nano body-stabilized LacY
dc.typeArticle
dc.date.updated2020-05-08T03:13:16Z
prism.issueIdentifier5
prism.publicationNamePLOS ONE
prism.volume15
dc.identifier.doi10.17863/CAM.52225
dcterms.dateAccepted2020-04-22
rioxxterms.versionofrecord10.1371/journal.pone.0232846
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/
datacite.contributor.supervisoreditor: van Veen, Hendrik W.
dc.contributor.orcidSteyaert, Jan [0000-0002-3825-874X]
dc.contributor.orcidStroud, Robert M. [0000-0003-2083-5665]
dc.identifier.eissn1932-6203
pubs.funder-project-idNational Institutes of Health (GM024485)
pubs.funder-project-idNational Institutes of Health (GM120043)
pubs.funder-project-idNational Science Foundation (MCB1747705)
pubs.funder-project-idINSTRUCT-ERIC (none)
pubs.funder-project-idSandler Foundation (PBBR)
pubs.funder-project-idUniversity of California Office of the President (MR-15-328599)


Files in this item

Thumbnail
Thumbnail
Thumbnail

This item appears in the following Collection(s)

Show simple item record

Attribution 4.0 International (CC BY 4.0)
Except where otherwise noted, this item's licence is described as Attribution 4.0 International (CC BY 4.0)