Center for Matrix Biology - Vanderbilt University and Medical Center

Charles R. Sanders, Ph.D.
Professor in the Department of Biochemistry

	Office Address:	Rm 5110C BIOSCI/MRBIII
	Mailing Address:	Rm 5110C BIOSCI/MRBIII Nashville, TN 37232-8725
	Lab address:	5110C MRB III
	Phone:	(615) 936-3756
	FAX:	615-936-6274
		email \| website

Research Summary

Membrane Protein Misfolding and Disease

A surprising number of diseases are caused by missense mutations in genes encoding membrane proteins that result in a loss of function due to the misfolding and mistrafficking of the protein. We seek to elucidate the molecular bases for misfolding-based disease by carrying out studies of purified membrane proteins under well-controlled conditions and by correlating results from these studies to the folding/misfolding of the same proteins in vivo and in vitro. We are pursuing several systems. The first involves human peripheral myelin protein 22 (PMP22) of the peripheral nervous system. This protein has 4 transmembrane segments and undergos mutation-prompted misfolding which results in the most common peripheral neuropathy, Charcot-Marie-Tooth Disease. A second project involves the G protein-coupled vasopressin V2 receptor, for which mutations result in defective folding and the diabetes insipidus. A third project involves characterization of the amyloid precursor protein, from which the beta amyloid protein is derived that is closely associated with Alzheimer's disease. A fourth project involves characterizing the interactions of a family of membrane proteins that modulate the function of voltage-gated potassium channels-- interactions that are closely linked both the cardiovascular and auditory dysfunction. Finally, we are also examining the prokaryotic integral membrane protein, diacylglycerol kinase (DAGK). DAGK is an ideal model system for studies of membrane protein misfolding because, like a number of disease-related membrane proteins, it is hyper-susceptible to misfolding as a result of point mutations at any of many different positions. For each of these sytems we are examining the energetics, kinetics, and structural characteristics associated with folding and misfolding. These studies employ a wide range of techniques spanning the territory between molecular biophysics/biochemistry to cellular/molecular biology. Ultimately, we hope to use the information derived from these studies to develop novel therapeutics based on the avoidance or correction of misfolding.

NMR-Based Membrane Protein Structural Determination

Solution NMR has lagged behind X-ray crystallography in its ability to solve the structures of complex integral membrane proteins. We are presently pursuing the structure of diacylglycerol kinase in detergent micelles (100 kDa aggregate molecular weight). As a 42 kDa homotrimer with 9 transmembrane segments, DAGK represents a target for NMR structural analysis which is much larger than any membrane protein for which a structure has been solved to date by NMR methods. Indeed, as a target for structural analysis, DAGK is similar in size and complexity to many members of the G protein coupled-receptor family, a supremely enticing group of structural targets. We recently completed assignment of DAGK's backbone resonances and are proceding to complete determination of its 3-D structure. This work paves the way for new projects in the Sanders lab. NMR projects are also under way for each of the other proteins mentioned above.

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Associated initiative(s):

Career opportunities:
Postdoctoral Position Available

Selected Publications

» Beel, AJ, Sanders, CR Substrate specificity of gamma-secretase and other intramembrane proteases Cell Mol Life Sci 2008. 65:1311-34

» Kang, C, Tian, C, S??nnichsen, FD, Smith, JA, Meiler, J, George, AL, Vanoye, CG, Kim, HJ, Sanders, CR Structure of KCNE1 and implications for how it modulates the KCNQ1 potassium channel Biochemistry 2008. 47:7999-8006

» Sanders, CR Visiting order on membrane proteins by using nanotechnology Proc Natl Acad Sci U S A 2007. 104:6502-3

» Smith, JA, Vanoye, CG, George, AL, Meiler, J, Sanders, CR Structural models for the KCNQ1 voltage-gated potassium channel Biochemistry 2007. 46:14141-52

» Tian, C, Vanoye, CG, Kang, C, Welch, RC, Kim, HJ, George, AL, Sanders, CR Preparation, functional characterization, and NMR studies of human KCNE1, a voltage-gated potassium channel accessory subunit associated with deafness and long QT syndrome Biochemistry 2007. 46:11459-72

» Mobley, CK, Myers, JK, Hadziselimovic, A, Ellis, CD, Sanders, CR Purification and initiation of structural characterization of human peripheral myelin protein 22, an integral membrane protein linked to peripheral neuropathies Biochemistry 2007. 46:11185-95

» Sanders, CR, S??nnichsen, F Solution NMR of membrane proteins: practice and challenges Magn Reson Chem 2006. 44 Spec No:S24-40

» Mi, D, Kim, HJ, Hadziselimovic, A, Sanders, CR Irreversible misfolding of diacylglycerol kinase is independent of aggregation and occurs prior to trimerization and membrane association Biochemistry 2006. 45:10072-84

» Tian, C, Karra, MD, Ellis, CD, Jacob, J, Oxenoid, K, S??nnichsen, F, Sanders, CR Membrane protein preparation for TROSY NMR screening Methods Enzymol 2005. 394:321-34

» Tian, C, Breyer, RM, Kim, HJ, Karra, MD, Friedman, DB, Karpay, A, Sanders, CR Solution NMR spectroscopy of the human vasopressin V2 receptor, a G protein-coupled receptor J Am Chem Soc 2005. 127:8010-1

» Sanders, Charles R, Myers, Jeffrey K Disease-related misassembly of membrane proteins Annu Rev Biophys Biomol Struct 2004. 33:25-51

» Oxenoid, Kirill, Kim, Hak Jun, Jacob, Jaison, S??nnichsen, Frank D, Sanders, Charles R NMR Assignments for a Helical 40 kDa Membrane Protein J Am Chem Soc 2004. 126:5048-9

» Nagy, Joanna K, Sanders, Charles R Destabilizing mutations promote membrane protein misfolding Biochemistry 2004. 43:19-25

» Sanders, Charles R, Kuhn Hoffmann, Amy, Gray, Don N, Keyes, Melvin H, Ellis, Charles D French swimwear for membrane proteins Chembiochem 2004. 5:423-6

» Myers, Jeffrey K, Beihoffer, Lauren A, Sanders, Charles R Phenotology of disease-linked proteins Hum Mutat 2004. 25:90-97

» Nagy, Joanna K, Sanders, Charles R A critical residue in the folding pathway of an integral membrane protein Biochemistry 2002. 41:9021-5

» Luchette, P.A., Prosser, R.S., and Sanders, C.R Oxygen as a paramagnetic probe of membrane protein structure by cysteine mutagenesis and 19f NMR spectroscopy J. American Chemical Society 2002. 124:1778-1781

» Gorzelle, Bonnie M, Hoffman, Amy Kuhn, Keyes, Melvin H, Gray, Don N, Ray, Dale G, Sanders, Charles R Amphipols can support the activity of a membrane enzyme J Am Chem Soc 2002. 124:11594-5

» Oxenoid, Kirill, S??nnichsen, Frank D, Sanders, Charles R Topology and secondary structure of the N-terminal domain of diacylglycerol kinase Biochemistry 2002. 41:12876-82

» Luchette, PA, Prosser, RS, Sanders, CR Oxygen as a paramagnetic probe of membrane protein structure by cysteine mutagenesis and (19)F NMR spectroscopy J Am Chem Soc 2002. 124:1778-81

» Nagy, J K, Lonzer, W L, Sanders, C R Kinetic study of folding and misfolding of diacylglycerol kinase in model membranes Biochemistry 2001. 40:8971-80

» Oxenoid, K, S??nnichsen, FD, Sanders, CR Conformationally specific misfolding of an integral membrane protein Biochemistry 2001. 40:5111-8

» Sanders, C R, Ismail-Beigi, F, McEnery, M W Mutations of peripheral myelin protein 22 result in defective trafficking through mechanisms which may be common to diseases involving tetraspan membrane proteins Biochemistry 2001. 40:9453-9

» Nagy, J K, Kuhn Hoffmann, A, Keyes, M H, Gray, D N, Oxenoid, K, Sanders, C R Use of amphipathic polymers to deliver a membrane protein to lipid bilayers FEBS Lett 2001. 501:115-20

» Sanders, C R, Nagy, J K Misfolding of membrane proteins in health and disease: the lady or the tiger Curr Opin Struct Biol 2000. 10:438-42

» Czerski, L.,Sanders, C.R Thiol modification of membrane proteins dependence upon protein site membrane disposition and reagent hydrophobicity FEBS Letters 2000. 472:225-229

» Czerski, L, Vinogradova, O, Sanders, CR NMR-Based amide hydrogen-deuterium exchange measurements for complex membrane proteins: development and critical evaluation J Magn Reson 2000. 142:111-9

» L. Czerski, O. Vinogradova, and C.R. Sanders NMR-based amide hydrogen-deuterium exchange measurements for complex membrane proteins: development and critical evaluation J. of Magnetic Resonance 2000. 142:111-119

» Sanders, C R, Oxenoid, K Customizing model membranes and samples for NMR spectroscopic studies of complex membrane proteins Biochim Biophys Acta 2000. 1508:129-45

» Czerski, L, Sanders, C R Functionality of a membrane protein in bicelles Anal Biochem 2000. 284:327-33

» Gorzelle, B M, Nagy, J K, Oxenoid, K, Lonzer, W L, Cafiso, D S, Sanders, C R Reconstitutive refolding of diacylglycerol kinase, an integral membrane protein Biochemistry 1999. 38:16373-82

» Sanders, C R, Prosser, R S Bicelles: a model membrane system for all seasons Structure 1998. 6:1227-34

» Vinogradova, O, S??nnichsen, F, Sanders, CR On choosing a detergent for solution NMR studies of membrane proteins J Biomol NMR 1998. 11:381-6

» Vinogradova, O, Carlin, C, Sonnichsen, F D, Sanders, C R A membrane setting for the sorting motifs present in the adenovirus E3-13.7 protein which down-regulates the epidermal growth factor receptor J Biol Chem 1998. 273:17343-50

» Vinogradova, O, Badola, P, Czerski, L, S??nnichsen, FD, Sanders, CR Escherichia coli diacylglycerol kinase: a case study in the application of solution NMR methods to an integral membrane protein Biophys J 1997. 72:2688-701

» Badola, P, Sanders, C R Escherichia coli diacylglycerol kinase is an evolutionarily optimized membrane enzyme and catalyzes direct phosphoryl transfer J Biol Chem 1997. 272:24176-82

» Sanders, C R, Landis, G C Reconstitution of membrane proteins into lipid-rich bilayered mixed micelles for NMR studies Biochemistry 1995. 34:4030-40

» Sanders, CR Qualitative comparison of the bilayer-associated structures of diacylglycerol and a fluorinated analog based upon oriented sample NMR data Chem Phys Lipids 1994. 72:41-57

» C. R. Sanders and G. C. Landis Facile Acquisition and Assignment of Oriented Sample NMR Spectra for Bilayer Surface-Associated Proteins Journal of the American Chemical Society 1994. 116:6470-6471

» C. R. Sanders, B. J. Hare, K. Howard, and James H. Prestegard Magnetically Oriented Phospholipid Micelles as a Tool for the Study of Membrane-Associated Molecules Progress in NMR Spectroscopy 1994. 26:421-444

» Smith, R L, O''Toole, J F, Maguire, M E, Sanders, C R Membrane topology of Escherichia coli diacylglycerol kinase J Bacteriol 1994. 176:5459-65

» Sanders, CR Solid state 13C NMR of unlabeled phosphatidylcholine bilayers: spectral assignments and measurement of carbon-phosphorus dipolar couplings and 13C chemical shift anisotropies Biophys J 1993. 64:171-81

» Sanders, CR, Schwonek, JP Simulation of NMR data from oriented membrane proteins: practical information for experimental design Biophys J 1993. 65:1460-9

» Hare, BJ, Sanders, CR, McIntyre, SE, Prestegard, JH Synthesis and characterization of a 13C-labeled alpha-mannosyl glycolipid analog from [13C]glucose Chem Phys Lipids 1993. 66:155-8

» Sanders, CR, Schaff, JE, Prestegard, JH Orientational behavior of phosphatidylcholine bilayers in the presence of aromatic amphiphiles and a magnetic field Biophys J 1993. 64:1069-80

» Sanders, C R, Schwonek, J P An approximate model and empirical energy function for solute interactions with a water-phosphatidylcholine interface Biophys J 1993. 65:1207-18

» Sanders, C R, Schwonek, J P Characterization of magnetically orientable bilayers in mixtures of dihexanoylphosphatidylcholine and dimyristoylphosphatidylcholine by solid-state NMR Biochemistry 1992. 31:8898-905

» Ikeda-Saito, M, Hori, H, Andersson, LA, Prince, RC, Pickering, IJ, George, GN, Sanders, CR, Lutz, RS, McKelvey, EJ, Mattera, R Coordination structure of the ferric heme iron in engineered distal histidine myoglobin mutants J Biol Chem 1992. 267:22843-52

» Sanders, CR, Prestegard, JH Magnetically orientable phospholipid bilayers containing small amounts of a bile salt analogue, CHAPSO Biophys J 1990. 58:447-60

» Sanders, C R, Tian, G C, Tsai, M D Mechanism of adenylate kinase. Is there a relationship between local substrate dynamics, local binding energy, and the catalytic mechanism Biochemistry 1989. 28:9028-43

» Sanders, CR, Tsai, MD Ligand-protein interactions via nuclear magnetic resonance of quadrupolar nuclei Methods Enzymol 1989. 177:317-33