Lopes, J.L.S. and Nobre, T.M. and Cilli, E.M. and Beltramini, L.M. and Araujo, A.P.U. and Wallace, Bonnie A. (2014) Deconstructing the DGAT1 enzyme: binding sites and substrate interactions. Biochimica et Biophysica Acta (BBA) - Biomembranes 1838 (12), pp. 3145-3152. ISSN 0005-2736.
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Abstract
Diacylglycerol 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.
Metadata
| Item Type: | Article |
|---|---|
| Keyword(s) / Subject(s): | Diacylglycerol acyltransferase, Enzyme catalysis, Peptide–lipid interaction, Langmuir monolayer, Synchrotron radiation circular dichroism (SRCD) spectroscopy, Triglyceride synthesis |
| School: | Birkbeck Faculties and Schools > Faculty of Science > School of Natural Sciences |
| Research Centres and Institutes: | Bioinformatics, Bloomsbury Centre for (Closed), Structural Molecular Biology, Institute of (ISMB) |
| Depositing User: | Administrator |
| Date Deposited: | 04 Sep 2014 13:04 |
| Last Modified: | 02 Aug 2023 17:12 |
| URI: | https://eprints.bbk.ac.uk/id/eprint/10451 |
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