English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

Dynamic nuclear polarization study of inhibitor binding to the M2(18-60) proton transporter from influenza A.

MPS-Authors
/persons/resource/persons206080

Andreas,  L. B.
Research Group of Solid State NMR Spectroscopy-2, MPI for Biophysical Chemistry, Max Planck Society;

Locator
There are no locators available
Fulltext (public)
There are no public fulltexts available
Supplementary Material (public)

2517751_Suppl.pdf
(Supplementary material), 647KB

Citation

Andreas, L. B., Barnes, A. B., Corzilius, B., Chou, J. J., Miller, E. A., Caporini, M., et al. (2013). Dynamic nuclear polarization study of inhibitor binding to the M2(18-60) proton transporter from influenza A. Biochemistry, 52(16), 2774-2782. doi:10.1021/bi400150x.


Cite as: http://hdl.handle.net/11858/00-001M-0000-002E-9249-B
Abstract
We demonstrate the use of dynamic nuclear polarization (DNP) to elucidate ligand binding to a membrane protein using dipolar recoupling magic angle spinning (MAS) NMR. In particular, we detect drug binding in the proton transporter M2(18-60) from influenza A using recoupling experiments at room temperature and with cryogenic DNP. The results indicate that the pore binding site of rimantadine is correlated with previously reported widespread chemical shift changes, suggesting functional binding in the pore. Futhermore, the N-15-labeled ammonium of rimantadine was observed near A30 C-13 beta and G34 C-13 alpha, suggesting a possible hydrogen bond to A30 carbonyl. Cryogenic DNP was required to observe the weaker external binding site(s) in a ZF-TEDOR spectrum. This approach is generally applicable, particularly for weakly bound ligands, in which case the application of MAS NMR dipolar recoupling requires the low temperatures to quench dynamic exchange processes. For the fully protonated samples investigated, we observed DNP signal enhancements of similar to 10 at 400 MHz using only 4-6 mM of the polarizing agent TOTAPOL. At 600 MHz and with DNP, we measured a distance between the drug and the protein to a precision of 0.2 angstrom.