Application | Comment | Organism |
---|---|---|
biofuel production | DGAT1 is a target for genetic manipulation to increase seed oil production in oleaginous plants. Triacylglycerol produced by the enzyme can be a petrochemical alternative | Brassica napus |
Cloned (Comment) | Organism |
---|---|
gene DGAT1, recombinant expression of N-terminally His-tagged enzyme cytoplasmic and autoinhibitory domains, BnaDGAT11-113 and BnaDGAT11-80, in Escherichia coli strain BL21(DE3), method optimization and evaluation, overview. The tag is located at the disordered N-terminal region, which is distal to the folded C-terminal region and the primary ligand binding site. Hence tag-induced secondary structure elements, if any, might not influence the ligand binding properties | Brassica napus |
Crystallization (Comment) | Organism |
---|---|
purified recombinnat truncated enzyme forms, BnaDGAT11-113 and BnaDGAT11-80, X-ray diffraction structure determination and analysis. Synchrotron small-angle X-ray scattering (SAXS) experiments are performed to obtain detailed 3D structural information on this disordered region of DGAT1, multi-state modeling | Brassica napus |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of truncated enzyme mutants, BnaDGAT11-113 comprises the full-length cytoplasmic domain, and BnaDGAT11-80 comprises the autoinhibitory domain, the truncated proteins are non-globular and monomeric in vitro | Brassica napus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
CoA | an allosteric modulator of DGAT1, interaction with the cytoplasmic N-terminal region, overview | Brassica napus | |
additional information | the cytoplasmic N-terminal domain of Brassica napus diacylglycerol acyltransferase, (DGAT1) includes an inhibitory module and allosteric binding sites, conformational heterogeneity in the N-terminal domain of DGAT1 | Brassica napus | |
oleoyl-CoA | an allosteric modulator of DGAT1, interaction with the cytoplasmic N-terminal region, overview | Brassica napus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
endoplasmic reticulum membrane | - |
Brassica napus | 5789 | - |
additional information | DGAT1 (BnaDGAT1) is a polytopic membrane protein in the endoplasmic reticulum (ER) with its N-terminal domain (residues 1-113) localized to the cytoplasm. Structure model of isozyme DGAT1-a transmembrane and cytosolic domains, overview | Brassica napus | - |
- |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
8600 | - |
recombinant His-tagged cytosolic domain BnaDGAT11-80, gel filtration | Brassica napus |
13900 | - |
recombinant His-tagged cytosolic domain BnaDGAT11-113, gel filtration | Brassica napus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
acyl-CoA + 1,2-diacyl-sn-glycerol | Brassica napus | - |
CoA + 1,2,3-triacylglycerol | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Brassica napus | K9LL63 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme cytoplasmic and autoinhibitory domains BnaDGAT11-113 and BnaDGAT11-80 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography. TEV protease tag cleavage as well as enzyme protein concentration after affinity chromatography lead to protein aggregation and are omitted | Brassica napus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acyl-CoA + 1,2-diacyl-sn-glycerol | - |
Brassica napus | CoA + 1,2,3-triacylglycerol | - |
? | |
additional information | interaction of CoA and oleoyl-CoA with the cytoplasmic N-terminal region (BnaDGAT11-113) of isoform BnaC.DGAT1.a. Truncated forms BnaDGAT11-113 and BnaDGAT11-80 interact with oleoyl-CoA or CoA with micromolar affinity, docking study and kinetics. The N-terminal domain of BnaDGAT1 has a higher affinity for thioester than free CoA. Interestingly, BnaDGAT11-80 also interacts with both ligands but with lower affinity. Ligand binding results in gain of secondary structure in mutants BnaDGAT11-113 and BnaDGAT11-80 | Brassica napus | ? | - |
- |
|
oleoyl-CoA + 1,2-diacyl-sn-glycerol | - |
Brassica napus | CoA + 1,2-diacyl-3-oleoyl-glycerol | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | 1 * 21000, about, recombinant His-tagged cytosolic domain BnaDGAT11-113, sequence calculation, 1 * 10500, about, recombinant His-tagged autoinhibitory domain BnaDGAT11-80, sequence calculation | Brassica napus |
Synonyms | Comment | Organism |
---|---|---|
BnaC.DGAT1.a | - |
Brassica napus |
DGAT1 | - |
Brassica napus |
diacylglycerol acyltransferase 1 | - |
Brassica napus |
General Information | Comment | Organism |
---|---|---|
additional information | determination of conformational heterogeneity in the N-terminal domain of DGAT1, disorder propensity for the full-length BnaDGAT11-501 sequence, overview. A small gain of secondary structure is induced by ligand binding. The cytoplasmic N-terminal domain of Brassica napus diacylglycerol acyltransferase, (DGAT1) includes an inhibitory module and allosteric binding sites | Brassica napus |
physiological function | intrinsic disorder in plant proteins has been reported to be essential for the stress response. An intrinsically disordered region (IDR) spanning the N-terminal cytosolic domain of the intramembrane enzyme diacylglycerol acyltransferase1 (DGAT1) from canola-type Brassica napus, has been identified. The IDR spans amino acid residues 1-80, while residues 81-113 have a folded structure. DGAT1 (EC 2.3.1.20) catalyzes the acyl-coenzyme A (CoA)-dependent acylation of sn-1, 2-diacylglycerol (DAG) to produce triacylglycerol (TAG) and CoA. TAG serves as an energy source for germination in plants, a component of edible oil. This enzyme has a substantial effect on carbon flux into seed oil | Brassica napus |