The primary goals of this competing renewal application are to define the structural requirements for selective and potent binding to the glutamate synaptic vesicular transporter (VGLUT) and to advance our understanding of the structural and functional roles of VGLUT. The long-range goals of this study are to: (a) develop pharmacophore and structural models of VGLUT that will aid the development of selective molecules that can regulate vesicular storage, uptake and release of glutamate and, (b) define the role and contribution of VGLUTs and related vesicular transporter proteins in vesicle function. The proposed strategy will optimize VGLUT inhibition and binding selectivity using results from our current project combined with novel chemical, computational, biochemical and pharmacological approaches. A concurrent strategy will seek to obtain a more global understanding of VGLUT structure and function. Using integrative strategies, we will increase our project activity and broaden our efforts to define the structural, functional and pharmacological roles of VGLUT. For the proposed grant period, we will address the following objectives:
Specific Aim 1 : Design and synthesize inhibitor structures that selectively bind VGLUT. Emphasis will be placed on developing inhibitors that satisfy the elements of a dynamic pharmacophore model, for example, a cyclic platform with regio- and stereochemically defined placement of acid and lipophilic substituents.
Specific Aim 2 : Test the activity of compounds prepared in SA 1 as inhibitors of VGLUT. The ability of these compounds to bind EAA receptors and cellular transporters also will be determined to assess the specificity at VGLUT.
Specific Aim 3 : Generate a computationally-derived pharmacophore model of VGLUT utilizing the chemical, biochemical and pharmacological data generated in SA's 1 & 2, The pharmacophore model will be used to refine and iterate inhibitor design (SA1), affinity agents (SA4), protein probe structures (SA's 4 and 5), and to create protein 3D structural hypotheses consistent with the probe and cross-linking results (SA4 & 5).
Specific Aim 4 : Characterize the transmembrane domain regions of VGLUT and neighboring architectures that contribute to VGLUT structure and function. Using N-term VGLUT antibodies, affinity tags, tethered- and cross-linking probes, VGLUT residues and structures proximal to VGLUT will be isolated and identified by mass spectrometry.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS038248-06
Application #
6881686
Study Section
Special Emphasis Panel (ZRG1-MDCN-4 (01))
Program Officer
Silberberg, Shai D
Project Start
1999-09-01
Project End
2007-03-30
Budget Start
2005-04-01
Budget End
2006-03-31
Support Year
6
Fiscal Year
2005
Total Cost
$332,500
Indirect Cost
Name
University of Montana
Department
Other Health Professions
Type
Schools of Pharmacy
DUNS #
010379790
City
Missoula
State
MT
Country
United States
Zip Code
59812
Stine, Jessica M; Ahl, Gabriel J H; Schlenker, Casey et al. (2017) The Interaction between the Third Type III Domain from Fibronectin and Anastellin Involves ?-Strand Exchange. Biochemistry 56:4667-4675
Carrigan, Christina N; Patel, Sarjubhai A; Cox, Holly D et al. (2014) The development of benzo- and naphtho-fused quinoline-2,4-dicarboxylic acids as vesicular glutamate transporter (VGLUT) inhibitors reveals a possible role for neuroactive steroids. Bioorg Med Chem Lett 24:850-4
Ahmed, S Kaleem; Etoga, Jean-Louis G; Patel, Sarjubhai A et al. (2011) Use of the hydantoin isostere to produce inhibitors showing selectivity toward the vesicular glutamate transporter versus the obligate exchange transporter system x(c)(-). Bioorg Med Chem Lett 21:4358-62
Etoga, Jean-Louis G; Ahmed, S Kaleem; Patel, Sarjubhai et al. (2010) Conformationally-restricted amino acid analogues bearing a distal sulfonic acid show selective inhibition of system x(c)(-) over the vesicular glutamate transporter. Bioorg Med Chem Lett 20:2680-3
Mullins, Jason E; Etoga, Jean-Louis G; Gajewski, Mariusz et al. (2009) Unexpected Formation of Highly Functionalized Dihydropyrans via Addition-Cyclization Reactions Between Dimethyl Oxoglutaconate and ?,?-Unsaturated Hydrazones. Tetrahedron Lett 50:2298-2300
Cox, Holly D; Thompson, Charles M (2008) Purification and proteomic analysis of synaptic vesicles. Methods Mol Biol 432:259-74
Cox, Holly D; Chao, Chih-Kai; Patel, Sarjubhai A et al. (2008) Efficient digestion and mass spectral analysis of vesicular glutamate transporter 1: a recombinant membrane protein expressed in yeast. J Proteome Res 7:570-8
Patel, Sarjubhai A; Nagy, Jon O; Bolstad, Erin D et al. (2007) Tetrapeptide inhibitors of the glutamate vesicular transporter (VGLUT). Bioorg Med Chem Lett 17:5125-8
Thompson, Charles M; Davis, Erin; Carrigan, Christina N et al. (2005) Inhibitor of the glutamate vesicular transporter (VGLUT). Curr Med Chem 12:2041-56
Bunik, Victoria I; Denton, Travis T; Xu, Hui et al. (2005) Phosphonate analogues of alpha-ketoglutarate inhibit the activity of the alpha-ketoglutarate dehydrogenase complex isolated from brain and in cultured cells. Biochemistry 44:10552-61

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