High-Sensitivity Detection of Nanometer 1H-19F Distances for Protein Structure Determination by 1H-Detected Fast MAS NMR

被引:32
作者
Shcherbakov, Alexander A. [1 ]
Mandala, Venkata Shiva [1 ]
Hong, Mei [1 ]
机构
[1] MIT, Dept Chem, 170 Albany St, Cambridge, MA 02139 USA
关键词
SOLID-STATE NMR; FULLY PROTONATED PROTEINS; ECHO DOUBLE-RESONANCE; ULTRAHIGH-RESOLUTION; ASSIGNMENT; BINDING; SPECTROSCOPY; BACKBONE; CHANNEL;
D O I
10.1021/acs.jpcb.9b03812
中图分类号
O64 [物理化学(理论化学)、化学物理学];
学科分类号
070304 ; 081704 ;
摘要
Protein structure determination by solid-state NMR requires the measurement of many interatomic distances through dipole-dipole couplings. To obtain multiple long-range distance restraints rapidly and with high sensitivity, here we demonstrate a new H-1-detected fast magic-angle-spinning NMR technique that yields many long distances in a two-dimensional (2D)-resolved fashion. The distances are measured up to similar to 15 angstrom, with an accuracy of better than 10%, between H-1 and F-19, two nuclear spins that have the highest gyromagnetic ratios. Exogenous fluorines are sparsely introduced into the aromatic residues of the protein, which is perdeuterated and back-exchanged to give amide protons. This H-1-F-19 distance experiment, termed 2D heteronuclear single-quantum coherence rotational-echo double-resonance (HSQC-REDOR), is demonstrated on the singly fluorinated model protein, GB1. We extracted 33 distances between 5-F-19-Trp43 and backbone amide protons, using 2D spectral series that were measured in less than 3 days. Combining these H-1-F-19 distance restraints with C-13-F-19 distances and chemical shifts, we calculated a GB1 structure with a backbone root-mean-square deviation of 1.73 angstrom from the high-resolution structure. This 1H detected H-1-F-19 distance technique promises to provide a highly efficient tool for constraining the three-dimensional structures of proteins and protein-ligand complexes, with not only precise and fast measurements but also access to truly long-range distances.
引用
收藏
页码:4387 / 4391
页数:5
相关论文
共 41 条
  • [31] 1H and 19F ultra-fast MAS double-quantum single-quantum NMR correlation experiments using three-spin terms of the dipolar homonuclear Hamiltonian
    Deschamps, Michael
    Fayon, Franck
    Cadars, Sylvian
    Rollet, Anne-Laure
    Massiot, Dominique
    [J]. PHYSICAL CHEMISTRY CHEMICAL PHYSICS, 2011, 13 (17) : 8024 - 8030
  • [32] Site-specific protein methyl deuterium quadrupolar patterns by proton-detected 3D H-2-C-13-H-1 MAS NMR spectroscopy
    Akbey, Umit
    [J]. JOURNAL OF BIOMOLECULAR NMR, 2022, 76 (1-2) : 23 - 28
  • [33] Sensitivity improvement of 1H-15N cross-polarization at high MAS frequency applied to NMR structural characterization of organic solids
    Palmas, Pascal
    Ilas, Simon
    Girard, Elzear
    [J]. MAGNETIC RESONANCE IN CHEMISTRY, 2011, 49 (12) : 788 - 794
  • [34] Probing Spatial Proximities Between Protons of Collagen Protein in Native Bone Using 2D 1H Multiple Quantum Experiments Under Fast MAS NMR
    Patra, Bijaylaxmi
    Agarwal, Vipin
    Nishiyama, Yusuke
    Sinha, Neeraj
    [J]. MAGNETIC RESONANCE IN CHEMISTRY, 2025, 63 (03) : 268 - 274
  • [35] Nano-Mole Scale Side-Chain Signal Assignment by 1H-Detected Protein Solid-State NMR by Ultra-Fast Magic-Angle Spinning and Stereo-Array Isotope Labeling
    Wang, Songlin
    Parthasarathy, Sudhakar
    Nishiyama, Yusuke
    Endo, Yuki
    Nemoto, Takahiro
    Yamauchi, Kazuo
    Asakura, Tetsuo
    Takeda, Mitsuhiro
    Terauchi, Tsutomu
    Kainosho, Masatsune
    Ishii, Yoshitaka
    [J]. PLOS ONE, 2015, 10 (04):
  • [36] 13C- and 1H-detection under fast MAS for the study of poorly available proteins: application to sub-milligram quantities of a 7 trans-membrane protein
    Dannatt, Hugh R. W.
    Taylor, Garrick F.
    Varga, Krisztina
    Higman, Victoria A.
    Pfeil, Marc-Philipp
    Asilmovska, Lubica
    Judge, Peter J.
    Watts, Anthony
    [J]. JOURNAL OF BIOMOLECULAR NMR, 2015, 62 (01) : 17 - 23
  • [37] 4′-C-[(4-Trifluoromethyl-1H-1,2,3-triazol-1-yl)methyl]thymidine as a Sensitive 19F NMR Sensor for the Detection of Oligonucleotide Secondary Structures
    Granqvist, Lotta
    Virta, Pasi
    [J]. JOURNAL OF ORGANIC CHEMISTRY, 2014, 79 (08) : 3529 - 3536
  • [38] 2H Chemical-shift resolution and dipolar 2H-1H, 2H-15N correlations in solid-state MAS NMR spectroscopy for structure determination and distance measurements in hydrogen-bonded systems
    Schulz-Dobrick, M
    Schnell, I
    [J]. CENTRAL EUROPEAN JOURNAL OF CHEMISTRY, 2005, 3 (02): : 245 - 251
  • [39] Accelerating the acquisition of high-resolution quadrupolar MQ/ST-HETCOR 2D spectra under fast MAS via 1H detection and through-space population transfers
    Sasaki, Akiko
    Trebosc, Julien
    Amoureux, Jean-Paul
    [J]. JOURNAL OF MAGNETIC RESONANCE, 2021, 333
  • [40] Average Intermolecular Distances and Dynamics of Ruthenium-Containing Organometallic Ionic Liquids Studied by Nuclear Overhauser Effects between 19F and 1H Nuclei and NMR Relaxation Rate Measurements
    Tei, Chikai
    Kuwahara, Daisuke
    Higashi, Tomomi
    Mochida, Tomoyuki
    [J]. BULLETIN OF THE CHEMICAL SOCIETY OF JAPAN, 2018, 91 (04) : 571 - 576