Klaus Gawrisch, PhD, Section Chief
National Institute on Alcohol Abuse and Alcoholism
National Institutes of Health
5625 Fishers Lane, Room 3N07:MSC 9410
Bethesda MD 20892-9410
telephone: +1 301 594 3750
fax: +1 301 594 0035
e-mail: klausg@mail.nih.gov
In addition to my responsibilities as Section Chief, I am conducting NMR structural studies on biomembranes by solid state NMR. My interests cover membrane biophysics, NMR structural studies on biomembranes, including G-protein coupled membrane receptors, structure and function of membranes rich in polyunsaturated fatty acids, lateral membrane organization, and the effects of ethanol on membrane structure and function.
Mission Statement
- Applies the techniques of low energy spectroscopy to the study of biomembrane architecture with respect to polyunsaturated phospholipids;
- defines microdomains with respect to phospholipid localization and molecular associations in the plasma membrane and their modification by ethanol;
- evaluates the feasibility and develop, as appropriate, NMR as an in vivo tool for the study of brain and liver desaturation and elongation of essential fatty acids;
- supports structure elucidation studies performed in other units of the laboratory; and
- collaborates closely on problems of mutual interest and using other biophysical techniques with others in the laboratory so that an interdisciplinary approach may be taken.
Current staff
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Alexei Yeliseev, PhD
Staff Scientist
301.443.0552
yeliseeva@mail.nih.gov
Current projects: My project focuses on developing methods for heterologous expression of functional cannabinoid receptor in large (milligram) quantities for NMR structural studies. Human peripheral-type cannabinoid receptor (CB2) is expressed in Escherichia coli as a fusion with the maltose-binding protein, solubilized in detergent micelles and purified by several steps of affinity chromatography. Functional activity and structural integrity of the receptor is confirmed by binding studies with a variety of natural and synthetic cannabinoid ligands and by activation of cognate G-proteins in an in vitro coupled assay. Reconstitution of the purified recombinant receptor into lipid matrix will generate samples suitable for structural studies by NMR and scattering techniques.
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Walter Teague, PhD
Chemist
301.443.3826
wteague@mail.nih.gov
Current projects: I am the NMR Facility Manager, in charge of maintenance and operation of three Nuclear Magnetic Resonance Spectrometers. I am carrying out computer system maintenance and assist in repair and diagnostics of all NMR equipment on site. Furthermore, I am responsible for the design and construction of unique pieces of scientific equipment used to assist in the study of reconstituted membranes containing GPCRs. My research is focused on biophysical properties of polyunsaturated lipid membranes.
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Olivier Soubias, PhD
Visiting Fellow
301.435.6727
soubiaso@mail.nih.gov
Current projects: My interests cover the development of NMR methods to study biomembranes, lipid-protein interaction, membrane organization, membrane biophysics and biochemistry. I am particularly interested in the interaction of polyunsaturated phospholipids with the G-protein coupled membrane receptors like rhodopsin. We utilized a novel reconstitution method for GPCR yielding single, tubular membranes in a porous anodic aluminum oxide support that yields very high volume concentrations of membranes. Saturation transfer MAS-NMR is applied to probe specific interactions between rhodopsin and lipids.
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Tomohiro Kimura, PhD
Visiting Fellow
301.594.3750
kimurato@mail.nih.gov
Current projects: I am attempting to elucidate the mechanisms of ligand binding to the G-protein coupled membrane receptors (GPCR) and the regulation of GPCR structure and function by the composition of the lipid matrix. I am reconstituting recombinant, purified human peripheral cannabinoid receptor CB2 into proteoliposomes with controlled membrane composition for structural and functional studies. Experiments include investigation of ligand binding to CB2 by solid- and solution-state NMR. Experiments utilize nanoporous anodized aluminium oxide (AAO) substrates for CB2 reconstitution.
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Lioudmila Zoubak, BS
Research Tech (Contractor)
301.443.0552
kimurato@mail.nih.gov
Current projects: I am working on expression of CB2 in E-coli bacteria. My responsibilities include tasks for modification of expression vectors, preparation of media, operation of equipment for large scale protein expression and purification, as well as ligand binding and G-protein activation studies using radioactive assays.
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Section Alumni
Nadukkudy. V Eldho, Ph.D., 2000-2003, Visiting Fellow: now Research Fellow, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH.
Holly C. Gaede, Ph.D., 2002-2005, Visiting Chemist: now Senior Lecturer and Assistant to the Associate Dean for Undergraduate Research, 2005 –Texas A&M University; College Station, TX
Daniel Huster, Ph.D. 1995-1999, Graduate Student: now Junior Professor, Head of Junior Research Group “Solid-state NMR studies of membrane-associated proteins”. Institute of Biotechnology, Martin Luther University Halle-Wittenberg, Germany
Laura L. Holte, Ph.D., 1993-1999, Postdoc IRTA: now NMR Scientist, Doty Scientific Inc. Columbia, SC
Bernd W. Koenig, Ph.D. 1996-1999, Visiting Fellow: now Staff Scientist, Forschungszentrum Juelich, Juelich, Germany
Dmitriy Krepkiy, Ph.D., 2004-2006,Visiting Fellow: now Research Fellow, NINDS, NIH
David T. Nizza, B.S., 1997-2001, Chemist: now Medical Resident, Reading Hospital, Medical Center, Reading PA, graduated from Medical School, University of Maryland, Baltimore, MD
Ivan V. Polozov, Ph.D., 1998-2004, Research Fellow: now Research Fellow, LCMB, NICHD, NIH
Wai-Ming Yau, Ph.D., 1996-2000, Postdoc IRTA: now Senior Research Assistant, Section Solid State NMR, LCP, NIDDK, NIH
Collaborators
Dr. V. Adrian Parsegian,: LPSB, NICHD, NIH
Dr. Ivan Polozov, Dr. Joshua Zimmerberg, , LCMB, NICHD, NIH
Dr. Scott E. Feller, Department of Chemistry, Wabash College, Crawfordsville, IN
Dr. Stephanie Tristram-Nagle, Dr. John F. Nagle, Dept. of Physics, Carnegie Mellon University, Pittsburgh, PA
Dr. Daniel Huster, Institute of Biotechnology, Martin Luther University Halle-Wittenberg, Germany
Dr. Sarah Veatch, Dr. Sarah Keller, Departments of Chemistry and Physics, University of Washington, Seattle, WA
Dr. Mihaela Mihailescu, Dr. Stephen H. White, Department of Physiology and Biophysics, School of Medicine, University of California, Irvine, CA
Dr. Susan Krueger, Dr. Charles Majkrzak, Cold Neutron Facility, NIST, Gaithersburg
Dr. Bernd Koenig, IBI-2, Research Center Juelich, Germany
Dr. Mike Pitman, IBM T. J. Watson Research Center, Yorktown Heights, NY
Selected Recent Publications
Gawrisch, K., Parsegian, V. A., Hajduk, D. A., Tate, M. W, Gruner, S. M., Fuller, N. L., Rand, R. P. (1992). Energetics of a hexagonal‑lamellar‑hexagonal‑phase transition sequence in dioleoylphosphatidylethanolamine membranes. Biochemistry 31: 2856-2864. PubMed
Gawrisch, K., Ruston, D., Zimmerberg, J., Parsegian, V. A., Rand, R. P., Fuller, N. (1992). Membrane dipole potentials, hydration forces and the ordering of water at membrane surfaces. Biophys. J. 61: 1213‑1223. PubMed
Gawrisch, K., Han, K.- H., Yang, J. - S., Bergelson, L. D., Ferretti, J. A. (1993). Interaction of a peptide fragment 828‑848 of the envelope glycoprotein of human immunodeficiency virus type I with lipid bilayers. Biochemistry 32: 3112-3118. PubMed
Holte, L. L., Peter, S. A., Sinnwell, T. M., Gawrisch, K. (1995). 2H NMR order parameter profiles suggest a change of molecular shape for phosphatidylcholines containing a polyunsaturated acyl chain. Biophys. J. 68: 2396-2403. PubMed
Holte, L. L., and Gawrisch, K. (1997). Determining ethanol distribution in phospholipid multilayers with MAS-NOESY spectra. Biochemistry 36: 4669-4674. PubMed
Koenig, B. W., Strey, H. H., Gawrisch, K. (1997). Membrane lateral compressibility measured by NMR and X-ray diffraction: Effect of acyl chain polyunsaturation. Biophysical J. 73: 1954-1966. PubMed
Yau, W.-M., Wimley, W. C., Gawrisch, K., White, S. H. (1998). The preference of tryptophan for membrane interfaces. Biochemistry 37: 14713-14718.
Huster, D., Arnold, K., Gawrisch, K. (1999). Investigation of lipid organization in biological membranes by two-dimensional Nuclear Overhauser Enhancement Spectroscopy. J. Phys. Chem. B 103: 243-251.
Huster, D. and Gawrisch, K. (1999). NOESY NMR crosspeaks between lipid headgroups and hydrocarbon chains: Spin diffusion or molecular disorder? J. Am. Chem. Soc. 121: 1992-1993.
Feller, S.E., Huster, D., Gawrisch, K. Interpretation of NOESY Cross-relaxation Rates from Molecular Dynamics Simulation of a Lipid Bilayer. J. Am. Chem. Soc. 121:8963-8964, 1999.
Yau, W.-M, Gawrisch, K. (2000) Lateral lipid diffusion dominates NOESY cross-relaxation in membranes. J. Am Chem. Soc. 122: 3971-3972.
Feller, S. E., Gawrisch, K., MacKerell, A. D. Jr. (2002) Polyunsaturated fatty acids in lipid bilayers: intrinsic and environmental contributions to their unique physical properties. J. Am. Chem. Soc. 124: 318-326. PubMed
Feller, S. E, Gawrisch, K, Woolf, T. B. (2003) Rhodopsin exhibits a preference for solvation by polyunsaturated docosahexaenoic acid. J. Am. Chem. Soc. 125: 4434-4435. PubMed
Eldho, N. V., Feller, S. E., Tristram-Nagle, S., Polozov, I. V., Gawrisch, K. (2003) Polyunsaturated Docosahexaenoic vs. Docosapentaenoic Acid – Differences in Lipid Matrix Properties from the Loss of One Double Bond. J. Am. Chem. Soc. 125: 6409-6421. PubMed
Gaede, H. C., Luckett, K. M., Polozov, I. V., Gawrisch, K. (2004) Multinuclear NMR studies of single lipid bilayers supported in cylindrical aluminum oxide nanopores. Langmuir 20: 7711-7719. PubMed
Gawrisch, K. (2005) The dynamics of membrane lipids. Chapter 4 in “The Structure of Biological Membranes”, second edition, ed. P.L. Yeagle, CRC Press, pp. 147-171.
Feller, S. E., Gawrisch, K. (2005) Properties of docosahexaenoic acid-containing lipids and their influence on the function of the GPCR rhodopsin. Current Opinion Struct. Biol. 15: 416-422. PubMed
Soubias, O., Gawrisch, K. (2005) Probing specific lipid-protein interaction by saturation transfer difference NMR spectroscopy. J. Am. Chem. Soc. 127: 13110-13111. PubMed
Yeliseev, A. A, Wong, K., Soubias, O., Gawrisch K. (2005) Expression of human peripheral cannabinoid receptor for structural studies. Protein Sci. 14: 2638-2653. PubMed
Polozov, I.V., Gawrisch, K. (2006) Characterization of the liquid-ordered state by proton MAS NMR. Biophys. J. 90:2051-2061. PubMed
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Last updated: June 2006
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