Activating Compound | Comment | Organism | Structure |
---|---|---|---|
16:0/16:0-phosphatidic acid | activated the enzyme by about 1.24fold | Brassica napus | |
16:0/18:1-phosphatidic acid | activates BnaDGAT1 by 2.4fold | Brassica napus | |
18:1/18:1-phosphatidic acid | activates BnaDGAT1 by 4.7fold | Brassica napus | |
phosphatidic acid | PA, Feed-forward activation of BnaDGAT1 by phosphatidate, PA facilitates the transition of the enzyme into the more active state. PA may also relieve possible autoinhibition of BnaDGAT1 brought about by the N-terminal regulatory domain, which is shown to interact with PA. The soluble domain of BnaDGAT1 is likely to interact with the small highly charged head group of PA. 18:1/18:1-PA and 16:0/18:1-PA activated BnaDGAT1 by about 4.7fold and 2.4fold, respectively. 16:0/16:0-PA only activates the enzyme by about 1.24fold, while 18:0/18:0-PA has a minimal effect on enzyme activity. Truncated enzyme BnaDGAT11-113, comprising the N-terminus, interacts with increasing amounts of PA, allowing the soluble domain to be recovered together with the liposomes upon centrifugation | Brassica napus |
Cloned (Comment) | Organism |
---|---|
gene DGAT1, BnaDGAT1 is recombinantly expressed using a codon-optimized cDNA in Saccharomyces cerevisiae strain H1246 microsomes, recombinant expression of truncated enzyme mutants in Saccharomyces cerevisiae strain H1246 microsomes | Brassica napus |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of truncated enzyme versions, BnaDGAT11-113 and BnaDGAT181-501. Yeast transformed with BnaDGAT181-501 with N-terminal Nub tag and SnRK1 with N-terminal Cub tag grew on selective media. Truncated enzyme BnaDGAT11-113, comprising the N-terminus, interacts with increasing amounts of PA, allowing the soluble domain to be recovered together with the liposomes upon centrifugation | Brassica napus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | phosphorylation downregulates the activity of the enzyme | Brassica napus | |
oleoyl-CoA | BnaDGAT1 exhibits a sigmoidal response and eventual substrate inhibition with respect to increasing concentrations of oleoyl-CoA. In the presence of phosphatidic acid (PA), the oleoyl-CoA saturation plot becomes more hyperbolic and desensitized to substrate inhibition. BnaDGAT1 is less susceptible to substrate inhibition at 0.005-0.020 mM oleoyl-CoA in the presence of 1 mg/ml BSA | Brassica napus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetic analysis of lipidated BnaDGAT1. BnaDGAT1 exhibits cooperative substrate binding behavior with oleoyl-CoA | Brassica napus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
microsome | - |
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 | Q9XGR5 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
phosphoprotein | BnaDGAT1 has a predicted SnRK1 phosphorylation site that appears to be conserved among plant DGAT1s. This site is localized in the cytosolic loop connecting the second and third predicted transmembrane segments in the TMHMM topology, indicating that it is accessible to cytoplasmic kinases. In vitro SnRK1-catalyzed phosphorylation of BnaDGAT1 results in a decrease in BnaDGAT1 activity | Brassica napus |
Purification (Comment) | Organism |
---|---|
recombinant BnaDGAT1 from Saccharomyces cerevisiae strain H1246 microsomes, purified Brassica napus diacylglycerol acyltransferase 1 (BnaDGAT1) in n-dodecyl-beta-D-maltopyranoside micelles is lipidated to form mixed micelles | Brassica napus |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
cell suspension culture | microspore-derived cell suspension cultures | 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 | relative increase in activity of microsomal BnaDGAT1, BnaDGAT181-501, and BnaDGAT2 at different concentrations of oleoyl-CoA upon addition of 18:1/18:1-phosphatidate in the reaction mixture. BnaDGAT1 exhibits a sigmoidal response and eventual substrate inhibition with respect to increasing concentrations of oleoyl-CoA, kinetics, overview | Brassica napus | ? | - |
- |
|
oleoyl-CoA + 1,2-diacyl-sn-glycerol | BnaDGAT1 exhibits cooperative substrate binding behavior with oleoyl-CoA. The lipidated BnaDGAT1 exhibited a sigmoidal response to increasing concentrations of oleoyl-CoA | Brassica napus | CoA + 1,2-diacyl-3-oleoyl-sn-glycerol | - |
? |
Synonyms | Comment | Organism |
---|---|---|
BnaDGAT1 | - |
Brassica napus |
diacylglycerol acyltransferase 1 | - |
Brassica napus |
General Information | Comment | Organism |
---|---|---|
metabolism | diacylglycerol acyltransferase 1 (DGAT1) catalyzes the final and committed step in the Kennedy pathway for triacylglycerol (TAG) biosynthesis | Brassica napus |
additional information | purified Brassica napus diacylglycerol acyltransferase 1 (BnaDGAT1) in n-dodecyl-beta-D-maltopyranoside micelles is lipidated to form mixed micelles. The degree of mixed micelle fluidity appears to influence acyltransferase activity. BnaDGAT1 exhibits a sigmoidal response and eventual substrate inhibition with respect to increasing concentrations of oleoyl-CoA. In the presence of phosphatidic acid (PA), the oleoyl-CoA saturation plot becomes more hyperbolic and desensitized to substrate inhibition indicating that PA facilitates the transition of the enzyme into the more active state. PA is a key effector modulating lipid homeostasis, in addition to its well recognized role in lipid signaling | Brassica napus |