Deconstructing the DGAT1 enzyme: Binding sites and substrate interactions

dc.contributorUniversidade Estadual Paulista (UNESP)
dc.creatorLopes, Jose L. S.
dc.creatorNobre, Thatyane M.
dc.creatorCilli, Eduardo M.
dc.creatorBeltramini, Leila M.
dc.creatorAraujo, Ana P. U.
dc.creatorWallace, B. A.
dc.date2015-03-18T15:53:19Z
dc.date2016-10-25T20:24:48Z
dc.date2015-03-18T15:53:19Z
dc.date2016-10-25T20:24:48Z
dc.date2014-12-01
dc.date.accessioned2017-04-06T07:02:39Z
dc.date.available2017-04-06T07:02:39Z
dc.identifierBiochimica Et Biophysica Acta-biomembranes. Amsterdam: Elsevier Science Bv, v. 1838, n. 12, p. 3145-3152, 2014.
dc.identifier0005-2736
dc.identifierhttp://hdl.handle.net/11449/116446
dc.identifierhttp://acervodigital.unesp.br/handle/11449/116446
dc.identifier10.1016/j.bbamem.2014.08.017
dc.identifierWOS:000343847200018
dc.identifier0000-0002-4767-0904
dc.identifierhttp://dx.doi.org/10.1016/j.bbamem.2014.08.017
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/927093
dc.descriptionDiacylglycerol 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.
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.languageeng
dc.publisherElsevier B.V.
dc.relationBiochimica Et Biophysica Acta-biomembranes
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.subjectDiacylglycerol acyltransferase
dc.subjectEnzyme catalysis
dc.subjectPeptide-lipid interaction
dc.subjectLangmuir monolayer
dc.subjectSynchrotron radiation circular dichroism (SRCD) spectroscopy
dc.subjectTriglyceride synthesis
dc.titleDeconstructing the DGAT1 enzyme: Binding sites and substrate interactions
dc.typeOtro


Este ítem pertenece a la siguiente institución