CORE--CHEMISTRY

Washington University, Saint Louis MO

Investigators

Linked publications & trials

Abstract

Numerous animal studies have demonstrated that the diabetic states induces a fatty acid-based metabolic dysfunction which has complex effects on multiple end-organ systems. Both accelerated atherosclerosis and diabetic cardiomyopathy are the major etiologic agents in the excessive cardiovascular mortality and morbidity present in diabetic patients. A central theme in diabetic cardiovascular disease is the dysfunctional accumulation of lipids in multiple critical cell types. The proposed research in the component projects focuses on the role of alterations in peroxisomal regulatory proteins (e.g., peroxisomal phospholipases) and proliferative elements (e.g., PPARalpha and PPARgamma) which accelerate atherosclerosis and predispose critical cells in the cardiovascular system to maladaptive metabolic alterations. Core A of the program project embodies five major technical advances either made in our early research or developed in the current chemistry Core. Provision of lipid-analytical, synthetic, and proteomic services to the component projects will facilitate the cost-effective accrual of vital scientific information for the elucidation of the molecular mechanisms underlying altered lipid metabolism in the diabetic state. The Core will offer a wide variety of state-of-the-art analytic (mass spectrometry, evaporative light scattering detection (ELSD), fluorescence spectrometry, quantitative PCR) and synthetic (chiral BEL, fluorescence probe for real-time PLA2 assays) expertise. Specifically Core A will perform: 1) analysis of polar and neutral lipid individual molecular species including free fatty acids, triacylglycerides, cholesterol, acylcarnitines, and all types of phospholipids from biological sources using state-of-the-art ESI/MS and ELSD with sensitivity at the subpicomole to picomole range; 2) proteins sequencing and protein post- translational modification determinations utilizing LC/MC (proteomics); 3) the synthesis of the iPA2 suicide inhibitor (i.e., BEL) and its resoled enantiomers which selectively inhibit iPA2beta dn iPLA2gamma; 4) real-time PLA2 activity assays utilizing novel fluorescence substrates we have developed; and 5) quantitative PCR utilizing Taqman methodology. As it has done during the current grant interval, the Core will continue to provide methodologic advances to facilitate the study of lipid and protein alterations in the diabetic state (see Progress Report).