Christian Griesinger

Max Planck Institute for Biophysical Chemistry, Göttingen, Lower Saxony, Germany

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Publications (407)2219.67 Total impact

  • Nilamoni Nath · Edward J d'Auvergne · Christian Griesinger
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    ABSTRACT: Together with NOE and J coupling, one-bond residual dipolar coupling (RDC), which reports on the three-dimensional orientation of an internuclear vector in the molecular frame, plays an important role in the conformation and configuration analysis of small molecules in solution by NMR spectroscopy. When the molecule has few CH bonds, or too many bonds are in parallel, the available RDCs may not be sufficient to obtain the alignment tensor used for structure elucidation. Long-range RDCs that connect nuclei over multiple bonds are normally not parallel to the single bonds and therefore complement one-bond RDCs. Herein we present a method for extracting the long-range RDC of a chosen proton or group of protons to all remotely connected carbon atoms, including non-protonated carbon atoms. Alignment tensors fitted directly to the total long-range couplings (T=J+D) enabled straightforward analysis of both the long-range and one-bond RDCs for strychnine.
    Angewandte Chemie International Edition 09/2015; DOI:10.1002/anie.201504432 · 11.26 Impact Factor
  • Marta G Carneiro · Jithender G Reddy · Christian Griesinger · Donghan Lee
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    ABSTRACT: Protein motions over various time scales are crucial for protein function. NMR relaxation dispersion experiments play a key role in explaining these motions. However, the study of slow conformational changes with lowly populated states remained elusive. The recently developed exchange-mediated saturation transfer experiments allow the detection and characterization of such motions, but require extensive measurement time. Here we show that, by making use of Fourier transform, the total acquisition time required to measure an exchange-mediated saturation transfer profile can be reduced by twofold in case that one applies linear prediction. In addition, we demonstrate that the analytical solution for R1ρ experiments can be used for fitting the exchange-mediated saturation transfer profile. Furthermore, we show that simultaneous analysis of exchange-mediated saturation transfer profiles with two different radio-frequency field strengths is required for accurate and precise characterization of the exchange process and the exchanging states.
    Journal of Biomolecular NMR 09/2015; DOI:10.1007/s10858-015-9985-9 · 3.14 Impact Factor
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    ABSTRACT: Prion diseases are fatal neurodegenerative diseases characterized by accumulation of the pathogenic prion protein PrP in the brain. We established quantitative real-time quaking-induced conversion for the measurement of minute amounts of PrP in body fluids such as urine. Using this approach, we monitored the efficacy of antiprion therapy by quantifying the seeding activity of PrP from the brain and urine of mice after prion infection. We found that the aggregation inhibitor anle138b decreased the levels of PrP in the brain and urine. Importantly, variations of PrP levels in the urine closely corresponded to those in the brain. Our findings indicate that quantification of urinary PrP enables measurement of prion disease progression in body fluids and can substitute for immunodetection in brain tissue. We expect PrP quantification biologic fluids (such as urine and cerebrospinal fluid) with quantitative real-time quaking-induced conversion to emerge as a valuable noninvasive diagnostic tool for monitoring disease progression and the efficacy of therapeutic approaches in animal studies and human clinical trials of prion diseases. Moreover, highly sensitive methods for quantifying pathologic aggregate seeds might provide novel molecular biomarkers for other neurodegenerative diseases that may involve prion-like mechanisms (protein aggregation and spreading), such as Alzheimer disease and Parkinson disease.
    08/2015; 74(9). DOI:10.1097/NEN.0000000000000233
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    ABSTRACT: INPHARMA (Internuclear NOEs for Pharmacophore Mapping) determines the relative orientation of two competitive ligands in the protein binding pocket. It is based on the observation of interligand transferred NOEs mediated by spin diffusion through protons of the protein and is, therefore, sensitive to the specific interactions of each of the two ligands with the protein. We show how this information can be directly included into a protein-ligand docking program to guide the prediction of the complex structures. Agreement between the experimental and back-calculated spectra based on the full relaxation matrix approach is translated into a score contribution that is combined with the scoring function ChemPLP of our docking tool PLANTS. This combined score is then used to predict the poses of 5 weakly bound cAMP-dependent Protein Kinase (PKA) ligands. After optimizing the setup, which finally also included trNOE data and optimized protonation states, very good success rates were obtained for all combinations of three ligands. For one additional ligand, no conclusive results could be obtained due to the ambiguous electron density of the ligand in the X-ray structure, which does not disprove alternative ligand poses. The failures of the remaining ligand are caused by suboptimal locations of specific protein side chains. Therefore, side-chain flexibility should be included in an improved INPHARMA-PLANTS version. This will reduce the strong dependence on the used protein input structure leading to improved scores not only for this last ligand.
    Journal of Chemical Information and Modeling 07/2015; DOI:10.1021/acs.jcim.5b00235 · 3.74 Impact Factor
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    ABSTRACT: Im Frühling 2013 versammelten sich NMR-Spektroskopiker am Weizmann-Institut in Israel, um neue Ansätze zur Verbesserung der Empfindlichkeit von NMR-Experimenten zu diskutieren, besonders im Hinblick auf Experimente an biomolekularen Systemen. Der vorliegende Aufsatz ist von vielen Autoren mit unterschiedlichem fachlichem Hintergrund verfasst worden; er beschreibt den Stand der Forschung auf diesem Gebiet, wie er im Rahmen des Treffens diskutiert wurde. Es werden Ansätze für Ultra-Hochfeld-NMR-Spektroskopie, für neuartige Techniken der NMR-Detektion, für Prinzipien und Methoden der Kernhyperpolarisation, aber auch für neue Probenpräparationsmethoden diskutiert. Diese Ansätze verbessern die Empfindlichkeit von NMR-Experimenten im Festkörper und in Lösung, sind voneinander unabhängig und können sich in ihren Auswirkungen multiplizieren. Auch wenn in allen genannten Bereichen bereits substanzielle Verbesserungen erzielt wurden, ist doch noch eine weite Strecke zu überwinden, um an die Empfindlichkeiten von optischen und elektronischen Spektroskopien heranzureichen. Diese Probleme und erste Lösungsansätze werden ebenfalls diskutiert.
    Angewandte Chemie 07/2015; DOI:10.1002/ange.201410653
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    ABSTRACT: In the Spring of 2013, NMR spectroscopists convened at the Weizmann Institute in Israel to brainstorm on approaches to improve the sensitivity of NMR experiments, particularly when applied in biomolecular settings. This multi-author interdisciplinary Review presents a state-of-the-art description of the primary approaches that were considered. Topics discussed included the future of ultrahigh-field NMR systems, emerging NMR detection technologies, new approaches to nuclear hyperpolarization, and progress in sample preparation. All of these are orthogonal efforts, whose gains could multiply and thereby enhance the sensitivity of solid- and liquid-state experiments. While substantial advances have been made in all these areas, numerous challenges remain in the quest of endowing NMR spectroscopy with the sensitivity that has characterized forms of spectroscopies based on electrical or optical measurements. These challenges, and the ways by which scientists and engineers are striving to solve them, are also addressed. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
    Angewandte Chemie International Edition 07/2015; 54(32). DOI:10.1002/anie.201410653 · 11.26 Impact Factor
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    ABSTRACT: Special diphenyl-pyrazole compounds and in particular anle138b, were found to reduce the progression of prion and Parkinson's disease in animal models. The therapeutic impact of these compounds was attributed to the modulation of α-synuclein and prion-protein aggregation related to these diseases. Photophysical and photochemical properties of the diphenyl-pyrazole compounds anle138b, anle186b and sery313b and their interaction with monomeric and aggregated α-synuclein were studied by fluorescence techniques. The fluorescence emission of diphenyl-pyrazole is strongly increased upon incubation with α-synuclein fibrils, while no change in fluorescence emission is found when brought in contact with monomeric α-synuclein. This points to a distinct interaction between diphenyl-pyrazole and the fibrillar structure with a high binding affinity (Kd = 190 ± 120 nM) for anle138b. Several α-synuclein proteins form a hydrophobic binding pocket for the diphenyl-pyrazole compound. A UV-induced dehalogenation reaction was observed for anle138b which is modulated by the hydrophobic environment of the fibrils. Fluorescence of the investigated diphenyl-pyrazole compounds strongly increases upon binding to fibrillar α-synuclein structures. Binding at high affinity occurs to hydrophobic pockets in the fibrils. The observed particular fluorescence properties of the dipenyl-pyrazole molecules open new possibilities for the investigation of the mode of action of these compounds in neurodegenerative diseases. The high binding affinity to aggregates and the strong increase in fluorescence upon binding make the compounds promising fluorescence markers for the analysis of aggregation-dependent epitopes. Copyright © 2015. Published by Elsevier B.V.
    Biochimica et Biophysica Acta 05/2015; 1850(9). DOI:10.1016/j.bbagen.2015.05.021 · 4.66 Impact Factor
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    ABSTRACT: Growing evidence supports a link between brain copper homeostasis, the formation of alpha-synuclein (AS)-copper complexes and the development of Parkinson´s disease (PD). Recently it was demonstrated that the physiological form of AS is N-terminally acetylated (AcAS). Here we used NMR spectroscopy to structurally characterize the interaction between Cu(I) and AcAS. We found that the formation of an AcAS-Cu(I) complex at the N-terminal re-gion stabilizes local conformations with α-helical secondary structure and restricted motility. Our work provides new evidence into the metallo-biology of PD and opens new lines of research as the formation of AcAS-Cu(I) complex might impact on AcAS membrane binding and aggregation.
    Journal of the American Chemical Society 05/2015; 137(20). DOI:10.1021/jacs.5b01911 · 12.11 Impact Factor
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    ABSTRACT: Structure-based drug design (SBDD) is a powerful and widely used approach to optimize affinity of drug candidates. With the recently introduced INPHARMA method, the binding mode of small molecules to their protein target can be characterized even if no spectroscopic information about the protein is known. Here, we show that the combination of the spin-diffusion-based NMR methods INPHARMA, trNOE, and STD results in an accurate scoring function for docking modes and therefore determination of protein-ligand complex structures. Applications are shown on the model system protein kinase A and the drug targets glycogen phosphorylase and soluble epoxide hydrolase (sEH). Multiplexing of several ligands improves the reliability of the scoring function further. The new score allows in the case of sEH detecting two binding modes of the ligand in its binding site, which was corroborated by X-ray analysis. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
    Angewandte Chemie International Edition 04/2015; 127(22). DOI:10.1002/anie.201500671 · 11.26 Impact Factor
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    ABSTRACT: Lectins from different sources have been shown to interfere with HIV infection by binding to the sugars of viral-envelope glycoproteins. Three-dimensional atomic structures of a number of HIV-inactivating lectins have been determined, both as free proteins and in glycan-bound forms. However, details on the mechanism of recognition and binding to sugars are elusive. Herein we focus on the anti-HIV lectin OAA from Oscillatoria agardhii: We show that in the absence of sugars in solution, both the sugar-free and sugar-bound protein conformations that were observed in the X-ray crystal structures exist as conformational substates. Our results suggest that glycan recognition occurs by conformational selection within the ground state; this model differs from the popular "excited-state" model. Our findings provide further insight into molecular recognition of the major receptor on the HIV virus by OAA. These details can potentially be used for the optimization and/or development of preventive anti-HIV therapeutics. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
    Angewandte Chemie International Edition 04/2015; 127(22). DOI:10.1002/anie.201500213 · 11.26 Impact Factor
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    ABSTRACT: Lectins from different sources are known to interfere with HIV infection. The anti-viral activity is mediated by binding to high mannose sugars present on the viral envelope, thereby inhibiting cell entry. The lectin from Oscillatoria agardhii agglutinin (OAA) specifically recognizes a unique substructure of high mannose sugars and exhibits broad anti-HIV activity. Here we report the assignment of backbone and side-chain (1)H, (13)C and (15)N resonances of free OAA.
    Biomolecular NMR Assignments 02/2015; 9(2). DOI:10.1007/s12104-015-9600-8 · 0.76 Impact Factor
  • Biophysical Journal 01/2015; 108(2):256a. DOI:10.1016/j.bpj.2014.11.1416 · 3.97 Impact Factor
  • Biophysical Journal 01/2015; 108(2):184a-185a. DOI:10.1016/j.bpj.2014.11.1020 · 3.97 Impact Factor
  • Roberto R. Gil · Christian Griesinger · Armando Navarro-Vázquez · Han Sun
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    ABSTRACT: The use of NMR spectroscopy in anisotropic media, particularly the application of residual dipolar couplings (RDCs) for the structural analysis of small molecules is presented. Experimental procedures such as choice and preparation of alignment media, as well as the selection of the right pulse-sequences is discussed along with computational techniques for data analysis. Methods for the analysis of flexible molecules are also presented and illustrated with several examples from literature.
    Structure Elucidation in Organic Chemistry, 12/2014: pages 279-324; , ISBN: 9783527333363
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    ABSTRACT: The eIF4E-binding protein 1 (4EBP1) has long been known to be completely unstructured without any secondary structures, which contributed significantly to the proposal of the induced fit mechanism for target binding of intrinsically disordered proteins. We show here that 4EBP1 is not completely unstructured, but contains a pre-structured helix.
    Molecular BioSystems 11/2014; 11(2). DOI:10.1039/C4MB00532E · 3.21 Impact Factor
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    ABSTRACT: The voltage-dependent anion channel (VDAC) is the most abundant protein of the outer mitochondrial membrane and constitutes the major pathway for the transport of ADP, ATP, and other metabolites. In this multidisciplinary study we combined solid-state NMR, electrophysiology, and molecular dynamics simulations, to study the structure of the human VDAC isoform 2 in a lipid bilayer environment. We find that the structure of hVDAC2 is similar to the structure of hVDAC1, in line with recent investigations on zfVDAC2. However, hVDAC2 appears to exhibit an increased conformational heterogeneity compared to hVDAC1 which is reflected in broader solid-state NMR spectra and less defined electrophysiological profiles.
    Journal of Biomolecular NMR 11/2014; 61(3-4). DOI:10.1007/s10858-014-9876-5 · 3.14 Impact Factor
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    ABSTRACT: Motions play a vital role in the functions of many proteins. Discrete conformational transitions to excited states, happening on timescales of hundreds of microseconds, have been extensively characterized. On the other hand, the dynamics of the ground state are widely unexplored. Newly developed high-power relaxation dispersion experiments allow the detection of motions up to a one-digit microsecond timescale. These experiments showed that side chains in the hydrophobic core as well as at protein-protein interaction surfaces of both ubiquitin and the third immunoglobulin binding domain of protein G move on the microsecond timescale. Both proteins exhibit plasticity to this microsecond motion through redistribution of the populations of their side-chain rotamers, which interconvert on the picosecond to nanosecond timescale, making it likely that this "population shuffling" process is a general mechanism.
    Angewandte Chemie International Edition in English 11/2014; 54(1). DOI:10.1002/anie.201408890 · 13.45 Impact Factor
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    ABSTRACT: Intrinsically disordered proteins (IDPs) are involved in a wide variety of physiological and pathological processes and are best described by ensembles of rapidly interconverting conformers. Using fast field cycling relaxation measurements we here show that the IDP alpha-synuclein as well as a variety of other IDPs undergoes slow reorientations at timescales comparable to folded proteins. The slow motions are not perturbed by mutations in alpha-synuclein, which are related to genetic forms of Parkinson's disease, and do not depend on secondary and tertiary structural propensities. Ensemble-based hydrodynamic calculations suggest that the timescale of the underlying correlated motion is largely determined by hydrodynamic coupling between locally rigid segments. Our study indicates that long-range correlated dynamics are an intrinsic property of IDPs and offers a general physical mechanism of correlated motions in highly flexible biomolecular systems.
    Journal of the American Chemical Society 10/2014; 136(46). DOI:10.1021/ja506820r · 12.11 Impact Factor
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    ABSTRACT: In a conformational selection scenario, manipulating the populations of binding-competent states should be expected to affect protein binding. We demonstrate how in silico designed point mutations within the core of ubiquitin, remote from the binding interface, change the binding specificity by shifting the conformational equilibrium of the ground-state ensemble between open and closed substates that have a similar population in the wild-type protein. Binding affinities determined by NMR titration experiments agree with the predictions, thereby showing that, indeed, a shift in the conformational equilibrium enables us to alter ubiquitin’s binding specificity and hence its function. Thus, we present a novel route towards designing specific binding by a conformational shift through exploiting the fact that conformational selection depends on the concentration of binding-competent substates.
    Angewandte Chemie 09/2014; 53(39):10367. DOI:10.1002/ange.201403102
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    ABSTRACT: The aggregation of alpha-synuclein (AS) is a critical step in the etiology of Parkinson's disease (PD) and other neurodegenerative synucleinopathies. This process is selectively enhanced by copper in vitro and the interaction is proposed to play a potential role in vivo. Presently, the identity of the Cu(I) binding sites in AS and their relative affinities are under debate. In this work we have addressed unresolved details related to the structural binding specificity and affinity of Cu(I) to full-length AS. We demonstrated conclusively that: (i) the binding preferences of Cu(I) for the Met-binding sites at the N- (K-d = 20 mu M) and C-terminus (K-d = 270 mu M) of AS are widely different: (ii) the imidazole ring of His-50 acts as an effective anchoring residue (K-d = 50 mu M) for Cu(I) binding to AS; and (iii) no major structural rearrangements occur in the protein upon Cu(I) binding. Overall, our work shows that Cu(I) binding to the N- and C-terminal regions of AS are two independent events, with substantial differences in their affinities, and suggest that protein oxidative damage derived from a misbalance in cellular copper homeostasis would target preferentially the N-terminal region of AS. This knowledge is key to understanding the structural-aggregation basis of the copper catalyzed oxidation of AS.
    Journal of Inorganic Biochemistry 09/2014; 141. DOI:10.1016/j.jinorgbio.2014.08.012 · 3.44 Impact Factor

Publication Stats

16k Citations
2,219.67 Total Impact Points


  • 2000–2015
    • Max Planck Institute for Biophysical Chemistry
      • Department of NMR-based Structural Biology
      Göttingen, Lower Saxony, Germany
  • 2014
    • Rosario National University
      Rosario, Santa Fe, Argentina
  • 1985–2012
    • Goethe-Universität Frankfurt am Main
      • • Institute of Biochemistry
      • • Institute of Organic Chemistry and Chemical Biology
      Frankfurt, Hesse, Germany
  • 2010
    • Paul-Ehrlich-Institut
      Langen, Hesse, Germany
  • 2001–2010
    • Massachusetts Institute of Technology
      • Department of Chemistry
      Cambridge, Massachusetts, United States
  • 2009
    • Universität Basel
      • Department of Biophysical Chemistry
      Bâle, Basel-City, Switzerland
  • 2008–2009
    • Max Planck Institute for Chemistry
      Mayence, Rheinland-Pfalz, Germany
    • Friedrich Schiller University Jena
      Jena, Thuringia, Germany
    • Technische Universität München
      München, Bavaria, Germany
    • Max Planck Research Unit for Structural Molecular Biology at DESY
      Hamburg, Hamburg, Germany
    • University of Alberta
      • Department of Biochemistry
      Edmonton, Alberta, Canada
    • European Molecular Biology Laboratory
      • Structural and Computational Biology Unit (Heidelberg)
      Heidelburg, Baden-Württemberg, Germany
    • University of Washington Seattle
      • Department of Biochemistry
      Seattle, Washington, United States
    • Texas A&M University
      • Department of Chemistry
      College Station, TX, United States
  • 2007
    • Max Planck Institute for Developmental Biology
      Tübingen, Baden-Württemberg, Germany
  • 1994–2007
    • University Hospital Frankfurt
      Frankfurt, Hesse, Germany
  • 2006
    • Imperial College London
      • Department of Chemistry
      London, ENG, United Kingdom
  • 2005
    • University of Bayreuth
      Bayreuth, Bavaria, Germany
  • 2003
    • Max Planck Institute of Biophysics
      Frankfurt, Hesse, Germany
  • 1995–2003
    • Philipps-Universität Marburg
      • Faculty of Biology
      Marburg, Hesse, Germany
  • 1996
    • Aarhus University
      • Department of Chemistry
      Aarhus, Central Jutland, Denmark
  • 1988–1991
    • Hochschule für Technik Zürich
      Zürich, Zurich, Switzerland
    • Max Planck Institute of Biochemistry
      • Department of Molecular Medicine
      München, Bavaria, Germany