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Protein-Strukturbiologie mit Kernmagnetresonanz-Spektroskopie (NMR)

Extraordinary advances occurred during the last few years in structural biology by nuclear magnetic resonance (NMR) spectroscopy. The precision and accuracy of solution structures by NMR improved dramatically and the upper range of molecular weights accessible to NMR structure determination is still increasing.

Once reasonable solution conditions are found, NMR structure determination is straight forward. Using nuclear spins in biomolecular structural studies has required exceptional efforts integrating chemical physics, instrumentation development, new algorithms, and labelling strategies.

NMR can provide a realistic, dynamic and/or time averaged view of a macromolecule ligand interface or, even, the transient intramolecular interfaces seen during protein folding not available by other methods. This is an advantage in probing the role of local entropic contributions to binding using relaxation methods, and in the dissection of localized weak contributions, as in the 'SAR (structure activity relationships) by NMR' approach.

Artikel 1

Strukturgalerie

c39


Lecher J, Schwarz CKW, Stoldt M, Smits SHJ, Willbold D, Schmitt L
RTX toxin transporters tether its substrate prior to secretion via the unique function of its N- terminal domain.
Structure 20, 1778-1787 (2012)

3ZUA

cnbd_apo

Schünke S, Stoldt M, Lecher J, Kaupp UB, Willbold D
Structural insights into conformational changes of a cyclic nucleotide-binding domain in solution from Mesorhizobium loti K1 channel.
Proc. Natl. Acad. Sci. USA 108, 6121-6126 (2011)

2KXL

atg8

Schwarten M, Stoldt M, Mohrlüder J, Willbold D
Solution structure of Atg8 reveals conformational polymorphism of the N-terminal domain.
Biochem. Biophys. Res. Comm. 395, 426-431 (2010)

2KQ7

cnbd_camp

Schünke S, Stoldt M, Novak K, Kaupp UB, Willbold D
Solution structure of the M.loti K1 channel cyclic nucleotide binding domain in complex with cAMP.
EMBO Rep. 10, 729-735 (2009)

2K0G

vpu

Wittlich M, Koenig BW, Stoldt M, Schmidt H, Willbold D
NMR structural characterization of HIV-1 virus protein U cytoplasmic domain in the presence of dodecylphosphatidylcholine micelles.
FEBS J. 276, 6560-75 (2009)

2K7Y

sh3

Schmidt H, Hoffmann S, Tran T, Stoldt M, Stangler T, Wiesehan K, Willbold D
Solution structure of a Hck SH3 domain ligand complex reveals novel interaction modes.
J. Mol. Biol. 365, 1517-1532 (2007)

2OI3

2OJ2

cd4

Wittlich M, Koenig BW, Hoffmann S, Willbold D
Structural characterisation of the transmembrane and cytoplasmic domains of human CD4.
Biochim Biophys Acta 1768, 2949-60 (2007)

2KLU

x4

Hänel K, Stangler T, Stoldt M, Willbold D
Solution structure of the X4 protein coded by the SARS related coronavirus reveals an immunoglobulin like fold and suggests a binding activity to integrin I domains.
J. Biomed. Sci. 13, 281-293 (2006)

1YO4

vpr

Engler A, Stangler T, Willbold D
Structure of human immunodeficiency virus type 1 Vpr (34-51) peptide in micelle containing aqueous solution.
Eur. J. Biochem. 269, 3264-3269 (2002)

1KZV

1KZT

1KZS

gabarap

Stangler T, Mayr LM, Willbold D
Solution structure of the GABAA receptor associated protein GABARAP: Implications for biological function and its regulation.
J. Biol. Chem. 277, 13363-13366 (2002)

1KOT



  • Aladag A, Hoffmann S, Stoldt M, Bösing C, Willbold D, Schwarten M
    Hepatitis C virus NS5A is able to competitively displace c-Myc from the Bin1 SH3 domain in vitro
    J. Pept. Science 20, 334-340 (2014)
  • Elter S, Raschle T, Arens S, Viegas A, Gelev V, Etzkorn M, Wagner G
    The use of amphipols for NMR structural characterization of 7-TM proteins
    J. Membr. Biol. in press, (2014)
    http://dx.doi.org/10.1007/s00232-014-9669-5
  • Etzkorn M, Zoonens M, Catoire LJ, Popot JL, Hiller S
    How Amphipols Embed Membrane Proteins: Global Solvent Accessibility and Interaction with a Flexible Protein Terminus
    J. Membr. Biol. in press, (2014)
    http://dx.doi.org/10.1007/s00232-014-9657-9
  • Grüning CS, Mirecka EA, Klein AN, Mandelkow E, Willbold D, Marino SF, Stoldt M, Hoyer W
    Alternative conformations of the tau repeat domain in complex with an engineered binding protein
    J. Biol. Chem. , (2014)
    http://dx.doi.org/10.1074/jbc.M114.560920
    http://www.jbc.org/content/early/2014/06/25/jbc.M114.560920.abstract
  • Ma P, Haller JD, Zajakala J, Macek P, Sivertsen AC, Willbold D, Boisbouvier J, Schanda P
    Probing transient conformational States of proteins by solid-state r1ρ relaxation-dispersion NMR spectroscopy.
    Angew. Chem. Int. Ed. Engl. 53, 4312–4317 (2014)
  • Mirecka EA, Shaykhalishahi H, Gauhar A, Akgül S, Lecher J, Willbold D, Stoldt M, Hoyer W
    Sequestration of a beta-hairpin for control of alpha-synuclein aggregation
    Angew. Chem. Int. Ed. 53, 4227-4230 (2014)
    http://dx.doi.org/10.1002/anie.201309001
    http://dx.doi.org/10.1002/ange.201309001
  • Bobby R, Medini K, Neudecker P, Lee TV, Brimble MA, McDonald FJ, Lott JS, Dingley AJ
    Structure and dynamics of human Nedd4-1 WW3 in complex with the αENaC PY motif
    Biochim Biophys Acta 1834, 1632-41 (2013)
    http://dx.doi.org/10.1016/j.bbapap.2013.04.031
  • Do H, Wittlich M, Glück JM, Möckel L, Willbold D, Koenig BW, Heise H
    Full-length Vpu and human CD4(372-433) in phospholipid bilayers as seen by magic angle spinning NMR.
    Biol. Chem. 394, 1453-1463 (2013)
  • Ma P, Schwarten M, Schneider L, Boeske A, Henke N, Lisak D, Weber S, Mohrlüder J, Stoldt M, Strodel B, Methner A, Hoffmann S, Weiergräber OH, Willbold D
    Interaction of Bcl-2 with the autophagy-related GABAA receptor-associated protein (GABARAP): Biophysical characterization and functional implications.
    J. Biol. Chem. 288, 37204-37215 (2013)
  • Rani R, Jentzsch K, Lecher L, Hartmann R, Willbold D, Jaeger K-E, Krauss U
    Conservation of dark recovery kinetic parameters and structural features in the Pseudomonadaceae ?short? LOV protein family ? implications for the design of LOV-based optogenetic tools.
    Biochemistry 52, 4460-4473 (2013)
  • Schwarten M, Solyom Z, Feuerstein S, Aladag A, Hoffmann S, Willbold D, Brutscher B
    Interaction of non-structural protein 5A of hepatitis C virus with SH3 domains using non-canonical binding sites
    Biochemistry 52, 6160-6168 (2013)
  • Schünke S, Stoldt M
    Structural snapshot of cyclic nucleotide binding domains from cyclic nucleotide-sensitive ion channels
    Biol Chem. 394, 1439-1451 (2013)
    http://www.ncbi.nlm.nih.gov/pubmed/24021595
  • Solyom Z, Schwarten M, Geist L, Konrat R, Willbold D, Brutscher B
    BEST-TROSY experiments for time-efficient sequential resonance assignment of large disordered proteins
    J. Biomol. NMR 55, 311-221 (2013)
  • Vasudevan AA, Smits SH, Hoeppner A, Haeussinger D, Koenig BW, Muenk C
    Structural features of antiviral DNA cytidine deaminases
    Biological Chemistry 394, 1357 - 1370 (2013)
  • Feuerstein S, Plevin MJ, Willbold D, Brutscher B
    iHADAMAC: a complementary tool for sequential resonance assignment of globular and highly disordered protein.
    J. Magn. Res. 214, 329-334 (2012)
  • Feuerstein S, Solyom Z, Aladag A, Favier A, Schwarten M, Hoffmann S, Willbold D, Brutscher B
    Transient structure and SH3 interaction sites in an intrinsically disordered fragment of the hepatitis C virus protein NS5A.
    J. Mol. Biol. 420, 310-323 (2012)
  • Holze R, Jacob T, Schünke S, Neudecker P, Willbold D
    Physical Chemistry 2011
    Nachr. Chemie 60, 313-322 (2012)
  • Koenig BW, Schünke S, Stoldt M, Willbold D
    NMR methods for the determination of protein-ligand interactions.
    Protein-Ligand Interactions" (Editor: H. Gohlke) (series "Methods and Principles in Medicinal Chemistry"), Wiley-VCH; ISBN: 3-527-32966-8, (2012)
  • Lecher J, Schwarz CKW, Stoldt M, Smits SHJ, Willbold D, Schmitt L
    RTX toxin transporters tether its substrate prior to secretion via the unique function of its N- terminal domain.
    Structure 20, 1778-1787 (2012)
  • Neudecker P, Robustelli P, Cavallli A, Walsh P, Lundstrom P, Zarrine-Afsar A, Sharpe S, Vendruscolo M, Kay LE
    Structure of an intermediate state in protein folding and aggregation.
    Science 336, 362-366 (2012)
  • Singh S, Möckel L, Thiagarajan-Rosenkranz P, Wittlich M, Willbold D, Koenig B
    Mapping the interaction between the cytoplasmic domains of HIV-1 VpU and human CD4 using NMR spectroscopy.
    FEBS J. 279, 3705-3714 (2012)
  • Sun N, Hartmann R, Lecher J, Stoldt M, Funke SA, Gremer L, Ludwig H-H, Demuth H-U, Kleinschmidt M, Willbold D
    Structural analysis of the pyroglutamate modified isoform of the Alzheimer's disease related beta-amyloid using NMR spectroscopy.
    J. Pept. Science 18, 691-695 (2012)
  • Wang W, Pacheco V, Krause H-J, Zhang Y, Dong H, Hartmann R, Willbold D, Offenhaeusser A, Gu Z
    Size and compositional effects on contrast efficiency of functionalized superparamagnetic nanoparticles at ultralow and ultrahigh magnetic fields.
    J. Phys. Chem. C 116, 17880-17884 (2012)
    10.1021/jp302758h
  • Feuerstein S, Solyom Z, Aladag A, Hoffmann S, Willbold D, Brutscher B
    1H, 13C, and 15N resonance assignment of a 179 residue fragment of hepatitis C virus nonstructural protein 5A
    Biomol. NMR Assign. 53, 241-243 (2011)
  • Glück JM
    Ligand interaction analysis of membrane-anchored proteins.
    Schriften des Forschungszentrums Jülich Reihe Gesundheit, Band 31, (2011)
    http://wwwzb1.fz-juelich.de/contentenrichment/inhaltsverzeichnisse/2010/verlag/gesund31_i.pdf
  • Lecher J, Stoldt M, Schwarz CKW, Smits SHJ, Schmitt L, Willbold D
    ¹H, ¹⁵N and ¹³C resonance assignment of the N-terminal C39 peptidase-like domain of the ABC transporter Haemolysin B (HlyB).
    Biomol. NMR Assign. 53, 199-201 (2011)
  • Schünke S, Stoldt M, Lecher J, Kaupp UB, Willbold D
    Structural insights into conformational changes of a cyclic nucleotide-binding domain in solution from Mesorhizobium loti K1 channel.
    Proc. Natl. Acad. Sci. USA 108, 6121-6126 (2011)
  • Seebahn A, Dinkel H, Mohrlüder J, Hartmann R, Vogel N, Becker CM, Sticht H, Enz R
    Structural characterization of intracellular C-terminal domains of group III metabotropic glutamate receptors
    FEBS Lett 585, 511-516 (2011)
  • Wang W, Dong H, Pacheco V, Willbold D, Zhang Y, Offenhausser A, Hartmann R, Weirich T, Ma P, Krause H, Gu Z
    Relaxation behavior study of ultra-small superparamagnetic iron oxide nanoparticles at ultra-low and ultra-high magnetic fields.
    J. Phys. Chem.B 115, 14789-14793 (2011)
  • Luheshi LM, Hoyer W, de Barros TP, van Dijk-Härd I, Brorsson AC, Macao B, Persson C, Crowther DC, Lomas DA, Ståhl S, Dobson CM, Härd T
    Sequestration of the Abeta peptide prevents toxicity and promotes degradation in vivo
    PLoS Biol. 8, e1000334 (2010)
  • Ma P, Mohrlüder J, Schwarten M, Stoldt M, Singh SK, Hartmann R, Pacheco V, Willbold D
    Preparation of a functional GABARAP-lipid conjugate in nanodiscs and its investigation by solution NMR spectroscopy
    Chembiochem 11, 1967-1970 (2010)
  • Pacheco V, Ma P, Thielmann Y, Hartmann R, Weiergräber OH, Mohrlüder J, Willbold D
    Assessment of GABARAP self-association by its diffusion properties
    J Biomol NMR 48, 49-58 (2010)
  • Schwarten M, Stoldt M, Mohrlüder J, Willbold D
    Solution structure of Atg8 reveals conformational polymorphism of the N-terminal domain
    Biochem Biophys Res Commun 395, 426-431 (2010)
  • Schünke S, Lecher J, Stoldt M, Kaupp UB, Willbold D
    Resonance assignments of the nucleotide-free wildtype MloK1 cyclic nucleotide-binding domain.
    Biomol. NMR Assign. 4, 147-150 (2010)
  • Smits SHJ, Meyer T, Mueller A, van Os N, Stoldt M, Willbold D, Schmitt L, Grieshaber MK
    Insights into the Mechanism of Ligand Binding to Octopine Dehydrogenase from Pecten maximus by NMR and Crystallography
    PLoS ONE 5, e12312 (2010)
    10.1371/journal.pone.0012312
    http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0012312
  • Wittlich M, Thiagarajan P, Koenig BW, Hartmann R, Willbold D
    NMR structure of the transmembrane and cytoplasmic domains of human CD4 in micelles.
    Biochim Biophys Acta 1798, 122-127 (2010)
  • Feuerstein SE, Pulvermüller A, Hartmann R, Granzin J, Stoldt M, Henklein P, Ernst OP, Heck M, Willbold D, Koenig BW
    Helix formation in Arrestin accompanies recognition of photoactivated Rhodopsin.
    Biochemsitry 48, 10733-42 (2009)
  • Glück JM, Wittlich M, Feuerstein S, Hoffmann S, Willbold D, Koenig BW
    Integral membrane proteins in nanodiscs can be studied by solution NMR spectroscopy.
    J. Am. Chem. Soc. 131, 12060-61 (2009)
  • Leliveld SR, Hendriks P, Michel M, Sajnani G, Bader V, Trossbach S, Prikulis I, Hartmann R, Jonas E, Willbold D, Requena JR, Korth C
    Oligomer assembly of the C-terminal DISC1 domain (640-854) is controlled by self-association motifs and disease-associated polymorphism S704C.
    Biochemistry 48, 7746-7755 (2009)
  • Mangels C, Frank AO, Ziegler J, Klingenstein R, Schweimer K, Willbold D, Korth C, Rösch P, Schwarzinger S
    Binding of TCA to the prion protein: mechanism, implication for therapy, and application as probe for complex formation of bio-macromolecules.
    J. Biomol. Struct. Dyn. 27, 163-170 (2009)
  • Schwarten M, Mohrlüder J, Ma P, Stoldt M, Thielmann Y, Stangler T, Hersch N, Hoffmann B, Merkel R, Willbold D
    Nix directly binds to GABARAP: a possible crosstalk between apoptosis and autophagy.
    Autophagy 5, 690-698 (2009)
  • Schwarten M, Stoldt M, Mohrlüder J, Willbold D
    Sequence-specific 1H, 13C, and 15N resonance assignment of the autophagy-related protein Atg8.
    Biomol NMR Assign 3, 137-139 (2009)
  • Schünke S, Stoldt M, Novak K, Kaupp UB, Willbold D
    Solution structure of the M.loti K1 channel cyclic nucleotide binding domain in complex with cAMP.
    EMBO Rep. 10, 729-735 (2009)
  • Thielmann Y, Weiergräber OH, Mohrlüder J, Willbold D
    Structural framework of the GABARAP-calreticulin interface - implications for substrate binding to endoplasmic reticulum chaperones
    FEBS J 276, 1140-1152 (2009)
  • Wittlich M, Koenig BW, Stoldt M, Schmidt H, Willbold D
    NMR structural characterization of HIV-1 virus protein U cytoplasmic domain in the presence of dodecylphosphatidylcholine micelles.
    FEBS J. 276, 6560-75 (2009)
  • Hartmann R, Koenig BW, Stangler T, Willbold D
    Structural Proteomics: Exploring Protein-Ligand Interactions by Solution NMR.
    in 'Proteomics Sample Preparation', J. v. Hagen, Wiley-VCH , 273-281 (2008)
  • Mueller CB, Loman A, Pacheco V, Koberling F, Willbold D, Richtering W, Enderlein J
    Precise measurement of diffusion by multi-color dual-focus fluorescence correlation spectroscopy.
    Europhys. Lett. 83, 46001 (2008)
  • Thielmann Y, Mohrlüder J, Koenig BW, Stangler T, Hartmann R, Becker K, Höltje HD, Willbold D
    An indole binding site is a major determinant of the ligand specificity of the GABA type A receptor-associated protein GABARAP
    Chembiochem 9, 1767-1775 (2008)
  • Weiergräber OH, Stangler T, Thielmann Y, Mohrlüder J, Wiesehan K, Willbold D
    Ligand binding mode of GABAA receptor-associated protein
    J Mol Biol 381, 1320-1331 (2008)
  • Wittlich M, Koenig BW, Willbold D
    Structural consequences of phosphorylation of two serine residues in the cytoplasmic domain of HIV-1 VpU.
    J. Pept. Sci. 14, 804-10 (2008)
  • Koenig BW
    Residual dipolar couplings report on the active conformation of rhodopsin-bound protein fragments.
    In: 'Topics in Current Chemistry', vol. 272, Bioactive Conformation I. T. Peters, editor. Springer-Verlag Berlin Heidelberg , 187-215 (2007)
  • Mohrlüder J, Hoffmann Y, Stangler T, Hänel K, Willbold D
    Identification of clathrin heavy chain as a direct interaction partner for the gamma-aminobutyric acid type A receptor associated protein
    Biochemistry 46, 14537-14543 (2007)
  • Mohrlüder J, Stangler T, Wiesehan K, Hoffmann Y, Mataruga A, Willbold D
    Identification of calreticulin as a ligand of GABARAP by phage display screening of a peptide library
    FEBS J 274, 5543-5555 (2007)
  • Scheidt HA, Vogel A, Eckhoff A, Koenig BW, Huster D
    Solid state NMR characterization of the putative membrane anchor of TWD1 from Arabidopsis thaliana.
    Eur. Biophys. J. 36, 393-404 (2007)
  • Schmidt H, Hoffmann S, Tran T, Stoldt M, Stangler T, Wiesehan K, Willbold D
    Solution structure of a Hck SH3 domain ligand complex reveals novel interaction modes.
    J. Mol. Biol. 365, 1517-1532 (2007)
  • Schünke S, Novak K, Stoldt M, Kaupp UB, Willbold D
    Resonance assignment of the cyclic nucleotide binding domain from a cyclic nucleotide-gated K+ channel in complex with cAMP.
    Biomol. NMR Assign. 1, 179-181 (2007)
  • Spiess HW, Jeschke G, Koenig BW, Willbold D
    Magnetische Resonanzspektroskopie.
    Nachr. Chemie 55, 288-293 (2007)
  • Stangler T, Hartmann R, Willbold D, Koenig BW
    Modern high resolution NMR for the study of structure, dynamics and interactions of biological macromolecules.
    In: 'Progress in Physical Chemistry', Vol. 1 / Different Aspects of Intermolecular Interaction, Oldenbourg, München , 111-157 (2007)
  • Stangler T, Tran T, Hoffmann S, Schmidt H, Jonas E, Willbold D
    Competitive displacement of full-length HIV-1 Nef from Hck SH3 domain by a high affinity artificial peptide.
    Biol. Chem. 388, 611-615 (2007)
  • Wittlich M, Koenig BW, Hoffmann S, Willbold D
    Structural characterisation of the transmembrane and cytoplasmic domains of human CD4.
    Biochim Biophys Acta 1768, 2949-60 (2007)
  • Batra-Safferling R, Abaraca-Heidemann K, Körschen HG, Tziatzios C, Stoldt M, Budyak I, Willbold D, Schwalbe H, Klein-Seetharaman J, Kaupp UB
    Glutamic acid-rich proteins of rod photoreceptors are natively unfolded.
    J Biol Chem 281, 1449-1460 (2006)
  • Hänel K, Stangler T, Stoldt M, Willbold D
    Solution structure of the X4 protein coded by the SARS related coronavirus reveals an immunoglobulin like fold and suggests a binding activity to integrin I domains.
    J. Biomed. Sci. 13, 281-293 (2006)
  • Stangler T, Hartmann R, Willbold D, Koenig BW
    Modern high resolution NMR for the study of structure, dynamics and interactions of biological macromolecules.
    Z. Phys. Chem. 220, 567-613 (2006)
  • Briese L, Preusser A, Willbold D
    Mapping the binding site of full length HIV-1 Nef on human Lck SH3 by NMR spectroscopy.
    J. Biomed. Sci. 12, 451-456 (2005)
  • Koenig BW, Gawrisch K
    Lipid ethanol interaction studied by NMR on bicelles.
    J. Phys. Chem. B. 109, 7540-47 (2005)
  • Niu AZ, Stellbrink J, Allgaier J, Willner L, Radulescu A, Richter D, Koenig BW, May RP, Fetters LJ
    An in situ study of the t-butyllithium initiated polymerization of butadiene in d-heptane via small angle neutron scattering and H-NMR.
    J. Chem. Phys. 122, 134906 (2005)
  • Willbold D
    Interaction of HIV-1 Nef with human CD4 and Lck.
    in 'Viral membrane proteins: structure, function and drug design', W. Fischer, ed. Kluwer Academic Publishers, New York , (2005)
  • Stoldt M, Kupce E, Rehm BHA, Görlach M
    Backbone resonance assignemnt of an aminogycoside-3'-phosphotransferase type IIa.
    J. Biomol. NMR 28, 93-94 (2004)
  • Willbold D, Koenig BW
    NMR Analysis of Biopolymer Structure.
    In 'Physics meets Biology: From Soft Matter to Cell Biology', G. Gompper, UB. Kaupp, JKG Dhont, D Richter, RG Winkler,eds. , B3.1-B3.14 (2004)
  • Briese L and Willbold D
    Structure determination of human Lck unique and SH3 domains by nuclear magentic resonance spectroscopy.
    BMC Struct. Biol. 3, 3 (2003)
  • Koenig BW, Rogowski M, Louis JM
    A rapid method to attain isotope labeled small soluble peptides for NMR studies.
    J. Biomol. NMR 26, 193-202 (2003)
  • Engler A, Stangler T, Willbold D
    Structure of human immunodeficiency virus type 1 Vpr (34-51) peptide in micelle containing aqueous solution.
    Eur. J. Biochem. 269, 3264-3269 (2002)
  • Gawrisch K, Koenig BW
    Lipid-peptide interactions investiagted by NMR.
    In: 'Current Topics in Membranes. Vol. 52: Peptide-lipid interactions' S.A. Simon and T. J. McIntosh, Eds., Academic Press, San Diego , 163-190 (2002)
  • Koenig BW
    Structure and orientation of ligands bound to membrane proteins are reflected by residual dipolar couplings in solution NMR experiments.
    ChemBioChem 3, 975-980 (2002)
  • Koenig BW, Kontaxis G, Mitchell DC, Louis JM, Litman BJ, Bax A
    Structure and orientation of a G protein fragment in the receptor bound state from residual couplings.
    J. Mol. Biol. 322, 441-461 (2002)
  • Stangler T, Mayr LM, Willbold D
    Solution structure of the GABA<SUB>A</SUB> receptor associated protein GABARAP: Implications for biological function and its regulation.
    J. Biol. Chem. 277, 13363-13366 (2002)
  • Briese L, Hoffmann S, Friedrich U, Biesinger B, Willbold D
    Sequence-specific 1H, 13C and 15N resonance assignments of lymphocyte specific kinase unique and SH3 domains.
    J. Biomol. NMR 19, 193-194 (2001)
  • Preusser A, Briese L, Baur AS, Willbold D
    Direct in vitro binding of full-length HIV-1 Nef protein to CD4 cytoplasmic domain.
    J. Virol. 75, 3960-3964 (2001)

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