Active Diacylglycerol-O-Acyltransferase Homolog 1 (DGAT1)

Diacylglycerol-O-Acyltransferase Homolog 1 (DGAT1) is a key enzyme located in the endoplasmic reticulum that catalyzes the final and committed step in triglyceride synthesis. It transfers a fatty acyl group from fatty acyl-CoA to diacylglycerol (DAG), forming a triglyceride. This reaction is essential for fat storage in adipose tissue and for the production of lipid droplets in various organs, playing a central role in overall energy homeostasis. DGAT1 works in conjunction with Monoacylglycerol-O-acyltransferase 2 (MOGAT2), which operates in the monoacylglycerol pathway. The functional interplay between DGAT1 and MOGAT2 is critical for efficient triglyceride synthesis and lipid metabolism, integrating two major metabolic pathways.To detect the activity of recombinant DGAT1, a functional ELISA assay was performed to evaluate the interaction between recombinant human DGAT1 and recombinant human MOGAT2.Briefly, biotin-linked DGAT1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100µl were then transferred to MOGAT2-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µl stop solution to the wells and read at 450nm immediately. The binding activity of human DGAT1 and recombinant human MOGAT2 was shown in Figure 1, and this effect was in a dose dependent manner.