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dc.contributor.authorVance, Stevenen
dc.contributor.authorTkachenko, Olgaen
dc.contributor.authorThomas, Benen
dc.contributor.authorBassuni, Monaen
dc.contributor.authorHong, Huien
dc.contributor.authorNietlispach, Danielen
dc.contributor.authorBroadhurst, Billen
dc.date.accessioned2016-02-23T16:20:39Z
dc.date.available2016-02-23T16:20:39Z
dc.date.issued2016-04-15en
dc.identifier.citationVance et al. Biochemical Journal (2016)en
dc.identifier.issn0264-6021
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/253979
dc.description.abstractType I modular polyketide synthases (PKSs) produce polyketide natural products by passing a growing acyl substrate chain between a series of enzyme domains housed within a gigantic multifunctional polypeptide assembly. Throughout each round of chain extension and modification reactions, the substrate stays covalently linked to an acyl carrier protein (ACP) domain. In the present study we report on the solution structure and dynamics of an ACP domain excised from MLSA2, module 9 of the PKS system that constructs the macrolactone ring of the toxin mycolactone, cause of the tropical disease Buruli ulcer. After modification of apo ACP with 4'-phosphopantetheine (Ppant) to create the holo form, (15)N nuclear spin relaxation and paramagnetic relaxation enhancement (PRE) experiments suggest that the prosthetic group swings freely. The minimal chemical shift perturbations displayed by Ppant-attached C3 and C4 acyl chains imply that these substrate-mimics remain exposed to solvent at the end of a flexible Ppant arm. By contrast, hexanoyl and octanoyl chains yield much larger chemical shift perturbations, indicating that they interact with the surface of the domain. The solution structure of octanoyl-ACP shows the Ppant arm bending to allow the acyl chain to nestle into a nonpolar pocket, whereas the prosthetic group itself remains largely solvent exposed. Although the highly reduced octanoyl group is not a natural substrate for the ACP from MLSA2, similar presentation modes would permit partner enzyme domains to recognize an acyl group while it is bound to the surface of its carrier protein, allowing simultaneous interactions with both the substrate and the ACP.
dc.description.sponsorshipThis work was supported by the Wellcome Trust [grant number 094252/Z/10/Z]. The Yousef Jameel Academic Foundation and the Cambridge Trust are thanked for providing a studentship to M.B.
dc.languageengen
dc.language.isoenen
dc.publisherPortland Press, Ltd.
dc.rightsAttribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subject4′-phosphopantetheineen
dc.subjectNMR spectroscopyen
dc.subjectacyl carrier proteinen
dc.subjectmycolactoneen
dc.subjecttype I polyketide synthaseen
dc.subjectAcyl Carrier Proteinen
dc.subjectMacrolidesen
dc.subjectMycobacterium ulceransen
dc.subjectPolyketide Synthasesen
dc.subjectProtein Structure, Secondaryen
dc.subjectProtein Structure, Tertiaryen
dc.titleSticky swinging arm dynamics: studies of an acyl carrier protein domain from the mycolactone polyketide synthase.en
dc.typeArticle
prism.endingPage1110
prism.issueIdentifier8en
prism.publicationDate2016en
prism.publicationNameBiochemical Journalen
prism.startingPage1097
prism.volume473en
dc.rioxxterms.funderWellcome Trust
dc.rioxxterms.projectid094252/Z10/Z
dcterms.dateAccepted2016-02-18en
rioxxterms.versionofrecord10.1042/BCJ20160041en
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/en
rioxxterms.licenseref.startdate2016-04-15en
dc.contributor.orcidNietlispach, Daniel [0000-0003-4364-9291]
dc.contributor.orcidBroadhurst, Bill [0000-0002-0264-4593]
dc.identifier.eissn1470-8728
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idWellcome Trust (094252/Z/10/Z)
pubs.funder-project-idBBSRC (BB/E013228/1)
cam.issuedOnline2016-02-26en


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