Deconstructing the DGAT1 enzyme: Binding sites and substrate interactions

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dc.contributor.authorLopes, Jose L. S.
dc.contributor.authorNobre, Thatyane M.
dc.contributor.authorCilli, Eduardo M. [UNESP]
dc.contributor.authorBeltramini, Leila M.
dc.contributor.authorAraujo, Ana P. U.
dc.contributor.authorWallace, B. A.
dc.contributor.institutionUniversidade de São Paulo (USP)
dc.contributor.institutionUniv London Birkbeck Coll
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.date.accessioned2015-03-18T15:53:19Z
dc.date.available2015-03-18T15:53:19Z
dc.date.issued2014-12-01
dc.description.abstractDiacylglycerol acyltransferase 1 (DGAT1) is a microsomal membrane enzyme responsible for the final step in the synthesis of triacylglycerides. Although DGATs from a wide range of organisms have nearly identical sequences, there is little structural information available for these enzymes. The substrate binding sites of DGAT1 are predicted to be in its large luminal extramembranous loop and to include common motifs with acyl-CoA cholesterol acyltransferase enzymes and the diacylglycerol binding domain found in protein kinases.In this study, synthetic peptides corresponding to the predicted binding sites of DGAT1 enzyme were examined using synchrotron radiation circular dichroism spectroscopy, fluorescence emission and adsorption onto lipid monolayers to determine their interactions with substrates associated with triacylglyceride synthesis (oleoyl-CoA and dioleoylglycerol). One of the peptides, Sit1, which includes the FYxDWWN motif common to both DGAT1 and acyl-CoA cholesterol acyltransferase, changes its conformation in the presence of both substrates, suggesting its capability to bind their acyl chains. The other peptide (Sit2), which includes the putative diacylglycerol binding domain HKWCIRHFYKP found in protein kinase C and diacylglycerol kinases, appears to interact with the charged headgroup region of the substrates. Moreover, in an extended-peptide which contains Sit1 and Sit2 sequences separated by a flexible linker, larger conformational changes were induced by both substrates, suggesting that the two binding sites may bring the substrates into close proximity within the membrane, thus catalyzing the formation of the triacylglyceride product. (C) 2014 Elsevier B.V. All Lights reserved.en
dc.description.affiliationUniv Sao Paulo, Inst Fis Sao Carlos, Sao Carlos, SP, Brazil
dc.description.affiliationUniv London Birkbeck Coll, Inst Struct & Mol Biol, London WC1E 7HX, England
dc.description.affiliationUniv Estadual Paulista, Inst Quim, Araraquara, Brazil
dc.description.affiliationUnespUniv Estadual Paulista, Inst Quim, Araraquara, Brazil
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipUK Biotechnology and Biological Sciences Research Council (BBSRC)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipBBSRC
dc.description.sponsorshipSRCD measurements at the ISA (Denmark), Soleil (France)
dc.description.sponsorshipANKA (Germany)
dc.description.sponsorshipEU Integrated Infrastructure Initiative
dc.description.sponsorshipEuropean Light Sources Activities (ELISA)
dc.description.sponsorshipIdFAPESP: 09/17698-5
dc.description.sponsorshipIdUK Biotechnology and Biological Sciences Research Council (BBSRC)H023852
dc.description.sponsorshipIdUK Biotechnology and Biological Sciences Research Council (BBSRC)J019135
dc.description.sponsorshipIdCNPq: 553056/2011-5
dc.description.sponsorshipIdCNPq: 407337/2013-0
dc.description.sponsorshipIdBBSRCJ019747
dc.description.sponsorshipIdEU Integrated Infrastructure Initiative13
dc.description.sponsorshipIdEuropean Light Sources Activities (ELISA)226716
dc.format.extent3145-3152
dc.identifierhttp://dx.doi.org/10.1016/j.bbamem.2014.08.017
dc.identifier.citationBiochimica Et Biophysica Acta-biomembranes. Amsterdam: Elsevier Science Bv, v. 1838, n. 12, p. 3145-3152, 2014.
dc.identifier.doi10.1016/j.bbamem.2014.08.017
dc.identifier.issn0005-2736
dc.identifier.orcid0000-0002-4767-0904
dc.identifier.urihttp://hdl.handle.net/11449/116446
dc.identifier.wosWOS:000343847200018
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofBiochimica Et Biophysica Acta-biomembranes
dc.relation.ispartofjcr3.438
dc.relation.ispartofsjr1,495
dc.rights.accessRightsAcesso restrito
dc.sourceWeb of Science
dc.subjectDiacylglycerol acyltransferaseen
dc.subjectEnzyme catalysisen
dc.subjectPeptide-lipid interactionen
dc.subjectLangmuir monolayeren
dc.subjectSynchrotron radiation circular dichroism (SRCD) spectroscopyen
dc.subjectTriglyceride synthesisen
dc.titleDeconstructing the DGAT1 enzyme: Binding sites and substrate interactionsen
dc.typeArtigo
dcterms.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dcterms.rightsHolderElsevier B.V.
unesp.author.orcid0000-0002-4767-0904[3]
unesp.campusUniversidade Estadual Paulista (Unesp), Instituto de Química, Araraquarapt

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