Unmasking crucial residues in adipose triglyceride lipase for coactivation with comparative gene identification-58.

Lipolysis is an essential metabolic process that releases unesterified fatty acids from neutral lipid stores to maintain energy homeostasis in living organisms. Adipose triglyceride lipase (ATGL) plays a key role in intracellular lipolysis and can be coactivated upon interaction with the protein comparative gene identification-58 (CGI-58). The underlying molecular mechanism of ATGL stimulation by CGI-58 is incompletely understood. Based on analysis of evolutionary conservation, we used site directed mutagenesis to study a C-terminally truncated variant and full-length mouse ATGL providing insights in the protein coactivation on a per-residue level. We identified the region from residues N209-N215 in ATGL as essential for coactivation by CGI-58. ATGL variants with amino acids exchanges in this region were still able to hydrolyze triacylglycerol at the basal level and to interact with CGI-58, yet could not be activated by CGI-58. Our studies also demonstrate that full-length mouse ATGL showed higher tolerance to specific single amino acid exchanges in the N209-N215 region upon CGI-58 coactivation compared to C-terminally truncated ATGL variants. The region is either directly involved in protein-protein interaction or essential for conformational changes required in the coactivation process. Three-dimensional models of the ATGL/CGI-58 complex with the artificial intelligence software AlphaFold demonstrated that a large surface area is involved in the protein-protein interaction. Mapping important amino acids for coactivation of both proteins, ATGL and CGI-58, onto the 3D model of the complex locates these essential amino acids at the predicted ATGL/CGI-58 interface thus strongly corroborating the significance of these residues in CGI-58-mediated coactivation of ATGL.

In vitro hypoglycaemic and hypolipidemic potential of white tea polyphenols.

The leaves at different processing stages of a single tea cultivar in order to obtain white (WT), green (GT) and black tea (BT) samples, were analysed. The capacities of tea polyphenolics to influence the glucose and lipid metabolism in HepG2 cell lines were evaluated. WT appeared the most active in reducing the glucose and cholesterol uptake (+17.7% and +32.4% in the glucose and cholesterol cell medium concentration, respectively). Incubation with WT enhanced LDL receptor binding activity by 40% (+20% for GT and +0% for BT) and led to an increase in HDL cell medium concentration of 33.3% (+20% for GT and +0% for BT). Finally, WT revealed the best inhibition capacity against lipase activity, and triglyceride levels in the cell medium increased by 400% (+382.6% for GT and +191.3% for BT). The present study intended to contribute to the little knowledge about the potential health benefits of white tea in individuals affected by metabolic syndrome.