Structural Determinants of Glycogen Synthase Regulation

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DESCRIPTION (provided by applicant): The overall goal of this proposal is to understand the structural and functional properties of the enzymes responsible for the synthesis and elongation of the glucose storage polymer, glycogen. Glycogen is an important storage reserve of glucose and the mechanism by which the synthesis of glycogen is initiated and elongated is conserved from yeast to humans. Our recent structural work on yeast glycogen synthase has provided important new insight into the functional properties of all eukaryotic enzymes. The long-range goal of this proposal is to build off this initial work to delineate the structural context for the reaction catalyzed by glycogen synthase and uncover the structural interplay between glycogen association and glucose-6- phosphate activation of glycogen synthase. As part of this effort, we will initialize a screen for small molecule modulators of human glycogen synthase that can manipulate the activity state of the enzyme and promote glucose disposal under conditions found in Type 2 Diabetes or inhibit the action of glycogen synthase as a potential approach for treating glycogen storage disorders, such as Lafora and Pompe's Disease. Structure determination will be an integral part or our efforts as complexes between glycogen synthase and its substrates in combination with glucose-6-phosphate or novel agonists/antagonists of the activator binding site will provide important structure/activity relationships that will guide further development of our research. As part of our screening efforts in Aim 2 we will produce novel enzymatic forms representing homogenous phosphorylation states and these will be tested against the novel modulators of glycogen synthase identified in this aim. In addition, we will pursue structure determination of these novel modulators in the presence and absence of substrates, substrate analogs or glucose-6-phosphate which will link to our work in aim 1. Aim 3 will determine the molecular mechanism of catalysis and the specificity of glycosyltransfer. One of the goals of this aim is to understand whether catalytic mistakes produced by glycogen synthase are ultimately responsible for aberrations in glycogen processing. This aim will draw upon the structural information developed in aim 1 and will employ wild-type and mutant forms of both yeast Gsy2p and human glycogen synthase and will feed important structure/activity information into our modulator discovery and development as part of aim 2.