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A yeast-based tool for screening mammalian diacylglycerol acyltransferase inhibitors
Peter Gajdoš1, Rodrigo Ledesma-Amaro2, Jean-Marc Nicaud3
1Institute of Biotechnology, Faculty of Chemical and Food Technology, Slovak University of Technology, Radlinskeho, Bratislava, Slovakia.
Abstract:
Dysregulation of lipid metabolism is associated with obesity and metabolic diseases but there is also increasing evidence of a relationship between lipid body excess and cancer. Lipid body synthesis requires diacylglycerol acyltransferases (DGATs) which catalyze the last step of triacylglycerol synthesis from diacylglycerol and acyl-coenzyme A. The DGATs and in particular DGAT2, are therefore considered potential therapeutic targets for the control of these pathologies. Here, the murine and the human DGAT2 were overexpressed in the oleaginous yeast Yarrowia lipolytica deleted for all DGAT activities, to evaluate the functionality of the enzymes in this heterologous host and DGAT activity inhibitors. This work provides evidence that mammalian DGATs expressed in Y. lipolytica are a useful tool for screening chemical libraries to identify potential inhibitors or activators of these enzymes of therapeutic interest.
Insights
Diacylglycerol acyltransferases (DGATs) are key in lipid metabolism and cancer. Mammalian DGAT2 enzymes expressed in yeast provide a new tool for discovering drugs targeting lipid-related diseases.
Area of Science:
- Biochemistry
- Molecular Biology
- Metabolic Diseases
Background:
- Lipid metabolism dysregulation is linked to obesity, metabolic diseases, and cancer.
- Diacylglycerol acyltransferases (DGATs) are crucial enzymes catalyzing the final step in triacylglycerol synthesis.
- DGATs, especially DGAT2, are potential therapeutic targets for these pathologies.
Purpose of the Study:
- To evaluate the functionality of murine and human DGAT2 in a heterologous yeast host (Yarrowia lipolytica).
- To establish a screening platform for DGAT activity inhibitors.
- To explore DGATs as therapeutic targets for metabolic diseases and cancer.
Main Methods:
- Overexpression of murine and human DGAT2 in Yarrowia lipolytica engineered for DGAT deficiency.
- Functional assessment of expressed mammalian DGAT2 enzymes in the yeast.
- Utilizing the engineered yeast strain for screening chemical libraries.
Main Results:
- Mammalian DGAT2 enzymes were successfully expressed and functional in the oleaginous yeast Yarrowia lipolytica.
- The engineered yeast system demonstrated utility in evaluating DGAT activity.
- This platform facilitates the identification of potential DGAT inhibitors or activators.
Conclusions:
- Yarrowia lipolytica is a suitable host for expressing functional mammalian DGAT2.
- Mammalian DGATs expressed in Y. lipolytica serve as a valuable tool for drug discovery.
- This approach aids in identifying therapeutic agents for diseases associated with lipid metabolism dysregulation.
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