Biochemistry of membrane proteins involved in lipid and cannabinoid metabolism

PROJECT SUMMARY The long-term goals of the Das lab are to identify the fundamental biochemical mechanisms underlying lipid metabolism and lipid-like xenobiotics such as cannabinoids. In the backdrop of growing recognition of lipids in the body and sparse knowledge of their biochemistry and function, the Das lab research program addresses the following two broad goals: (1) to understand the mechanism of how membrane proteins metabolize lipids and lipid-like xenobiotics, and (2) to identify novel lipid metabolites that are formed from the metabolism and elucidate their pharmacology with respect to receptor interactions that are involved in immunomodulation. To execute these research goals, Das lab uses diverse set of innovative methodologies spanning from chemical synthesis, recombinant membrane protein expression, nanodiscs, targeted and untargeted lipidomics mass spectrometry, UV-vis and fluorescence spectroscopy, molecular dynamics, imaging platforms, mammalian cell culture and receptor pharmacological studies. The project falls under the broader NIGMS mission as the elucidation of the basic mechanisms of membrane protein interactions with lipids and lipid-like xenobiotics will lay the foundation for the development of therapeutics for different diseases. The laboratory established in 2012 has been NIGMS funded and have published 52 papers from the independent laboratory focusing on CYP mediated metabolism of lipids. The work was recognized by four national awards and numerous opportunities to deliver talks at conferences and other academic institutions. Previously, the Das lab have combined use of cross-disciplinary approaches to understand the role of the lipid endocannabinoid metabolism by membrane-bound enzymes to synthesize novel metabolites that effect neuroinflammation. Overall, the laudable quality of our research program has been identification of important scientific problems, proposal of diverse solutions and finding collaborators to translate and strengthen the research question and approach. Using the tools and approaches developed in previous NIGMS funding, we will leverage support from the MIRA to understand the biochemistry of new orphan cytochrome P450s expressed during diseases states and involved in lipid metabolism. The strength in our studies lies in the central hypothesis that P450 mediated metabolism of both lipids and cannabinoids leads to the formation of bioactive molecules that are promising scaffold to develop therapeutics to treat immune diseases. Altogether, the proposal research will comprehensively address the mechanism of lipid membrane protein interactions leading to the formation of large repertoire of lipid metabolites that are involved in controlling systemic inflammation.