Real-time tracking of drug binding to influenza A M2 reveals a high energy barrier

被引:6
作者
Movellan, Kumar Tekwani [1 ]
Wegstroth, Melanie [1 ]
Overkamp, Kerstin [1 ]
Leonov, Andrei [1 ]
Becker, Stefan [1 ]
Andreas, Loren B. [1 ]
机构
[1] Max Planck Inst Multidisciplinary Sci, Dept NMR Based Struct Biol, Am Fassberg 11, Gottingen, Germany
关键词
Magic-angle spinning; Proton channel; Drug binding; Solid -state NMR; Binding kinetics; PROTON CHANNEL; VIRUS M2; LIPID-BILAYERS; HYDROGEN-BONDS; ION-CHANNEL; WILD-TYPE; AMANTADINE; MECHANISM; MEMBRANE; PROTEIN;
D O I
10.1016/j.yjsbx.2023.100090
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The drug Rimantadine binds to two different sites in the M2 protein from influenza A, a peripheral site and a pore site that is the primary site of efficacy. It remained enigmatic that pore binding did not occur in certain detergent micelles, and in particular incomplete binding was observed in a mixture of lipids selected to match the viral membrane. Here we show that two effects are responsible, namely changes in the protein upon pore binding that prevented detergent solubilization, and slow binding kinetics in the lipid samples. Using 55-100 kHz magic-angle spinning NMR, we characterize kinetics of drug binding in three different lipid environments: DPhPC, DPhPC with cholesterol and viral mimetic membrane lipid bilayers. Slow pharmacological binding kinetics allowed the characterization of spectral changes associated with non-specific binding to the protein periphery in the kinetically trapped pore-apo state. Resonance assignments were determined from a set of proton-detected 3D spectra. Chemical shift changes associated with functional binding in the pore of M2 were tracked in real time in order to estimate the activation energy. The binding kinetics are affected by pH and the lipid environment and in particular cholesterol. We found that the imidazole-imidazole hydrogen bond at residue histidine 37 is a stable feature of the protein across several lipid compositions. Pore binding breaks the imidazole-imidazole hydrogen bond and limits solubilization in DHPC detergent.
引用
收藏
页数:9
相关论文
共 55 条
[1]   Structure and mechanism of proton transport through the transmembrane tetrameric M2 protein bundle of the influenza A virus [J].
Acharya, Rudresh ;
Carnevale, Vincenzo ;
Fiorin, Giacomo ;
Levine, Benjamin G. ;
Polishchuk, Alexei L. ;
Balannik, Victoria ;
Samish, Ilan ;
Lamb, Robert A. ;
Pinto, Lawrence H. ;
DeGrado, William F. ;
Klein, Michael L. .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2010, 107 (34) :15075-15080
[2]   Structure and Mechanism of the Influenza A M218-60 Dimer of Dimers [J].
Andreas, Loren B. ;
Reese, Marcel ;
Eddy, Matthew T. ;
Gelev, Vladimir ;
Ni, Qing Zhe ;
Miller, Eric A. ;
Emsley, Lyndon ;
Pintacuda, Guido ;
Chou, James J. ;
Griffin, Robert G. .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2015, 137 (47) :14877-14886
[3]   Dynamic Nuclear Polarization Study of Inhibitor Binding to the M218-60 Proton Transporter from Influenza A [J].
Andreas, Loren B. ;
Barnes, Alexander B. ;
Corzilius, Bjoern ;
Chou, James J. ;
Miller, Eric A. ;
Caporini, Marc ;
Rosay, Melanie ;
Griffin, Robert G. .
BIOCHEMISTRY, 2013, 52 (16) :2774-2782
[4]   Magic Angle Spinning NMR Investigation of Influenza A M218-60: Support for an Allosteric Mechanism of Inhibition [J].
Andreas, Loren B. ;
Eddy, Matthew T. ;
Pielak, Rafal M. ;
Chou, James ;
Griffin, Robert G. .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2010, 132 (32) :10958-10960
[5]   Design and Pharmacological Characterization of Inhibitors of Amantadine-Resistant Mutants of the M2 Ion Channel of Influenza A Virus [J].
Balannik, Victoria ;
Wang, Jun ;
Ohigashi, Yuki ;
Jing, Xianghong ;
Magavern, Emma ;
Lamb, Robert A. ;
DeGrado, William F. ;
Pinto, Lawrence H. .
BIOCHEMISTRY, 2009, 48 (50) :11872-11882
[6]   Rapid Proton-Detected NMR Assignment for Proteins with Fast Magic Angle Spinning [J].
Barbet-Massin, Emeline ;
Pell, Andrew J. ;
Retel, Joren S. ;
Andreas, Loren B. ;
Jaudzems, Kristaps ;
Franks, W. Trent ;
Nieuwkoop, Andrew J. ;
Hiller, Matthias ;
Higman, Victoria ;
Guerry, Paul ;
Bertarello, Andrea ;
Knight, Michael J. ;
Felletti, Michele ;
Le Marchand, Tanguy ;
Kotelovica, Svetlana ;
Akopjana, Inara ;
Tars, Kaspars ;
Stoppini, Monica ;
Bellotti, Vittorio ;
Bolognesi, Martino ;
Ricagno, Stefano ;
Chou, James J. ;
Griffin, Robert G. ;
Oschkinat, Hartmut ;
Lesage, Anne ;
Emsley, Lyndon ;
Herrmann, Torsten ;
Pintacuda, Guido .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2014, 136 (35) :12489-12497
[7]   Membrane-Dependent Effects of a Cytoplasmic Helix on the Structure and Drug Binding of the Influenza Virus M2 Protein [J].
Cady, Sarah ;
Wang, Tuo ;
Hong, Mei .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2011, 133 (30) :11572-11579
[8]   Amantadine-induced conformational and dynamical changes of the influenza M2 transmembrane proton channel [J].
Cady, Sarah D. ;
Hong, Mei .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2008, 105 (05) :1483-1488
[9]   Structure of the amantadine binding site of influenza M2 proton channels in lipid bilayers [J].
Cady, Sarah D. ;
Schmidt-Rohr, Klaus ;
Wang, Jun ;
Soto, Cinque S. ;
DeGrado, William F. ;
Hong, Mei .
NATURE, 2010, 463 (7281) :689-U127
[10]   Magic Angle Spinning and Oriented Sample Solid-State NMR Structural Restraints Combine for Influenza A M2 Protein Functional Insights [J].
Can, Thach V. ;
Sharma, Mukesh ;
Hung, Ivan ;
Gor'kov, Peter L. ;
Brey, William W. ;
Cross, Timothy A. .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2012, 134 (22) :9022-9025