Analysis of influenza A virus NS1 dimer interfaces in solution by pulse EPR distance measurements
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Pulsed electron-electron double resonance (PELDOR) is an electron paramagnetic resonance (EPR) spectroscopy technique for nanometer distance measurements between paramagnetic centres such as radicals. PELDOR has been recognized as a valuable tool to approach structural questions in biological systems. In this manuscript we demonstrate the value of distance measurements for differentiating competing structural models on the dimerization of the effector domain (ED) of the non-structural protein 1 (NS1) of the influenza A virus. Our results show NS1 to be well amenable to nanometer distance measurements by EPR, yielding high quality data. In combination with mutants perturbing protein dimerization and in silico prediction based on crystal structures we can exclude one of two potential dimerization interfaces. Furthermore, our results lead to a viable hypothesis of a NS1 ED:ED interface which is flexible through rotation around the vector interconnecting the two native cysteines. These results prove the high value of pulse EPR as a complementary method for structural biology.
Kerry , P S , Turkington , H L , Ackermann , K , Jameison , S A & Bode , B E 2014 , ' Analysis of influenza A virus NS1 dimer interfaces in solution by pulse EPR distance measurements ' Journal of Physical Chemistry B , vol 118 , no. 37 , pp. 10882-10888 . DOI: 10.1021/jp508386r
Journal of Physical Chemistry B
© 2014 American Chemical Society. This is an Open Access article distributed in accordance with the terms of the Creative Commons Attribution (CC-BY) license, which permits others to distribute, remix, adapt and build upon this work, for commercial use, provided the original work is properly cited
H.L.T. was supported by the University of St Andrews; S.A.J. is supported by the BBSRC, U.K.; P.S.K. is supported by the Medical Research Council, U.K.; B.E.B. is grateful for an EaStCHEM Hirst Academic Fellowship by the School of Chemistry, St Andrews, and funding from the People Programme (Marie Curie Actions) of the European Union’s Seventh Framework Programme [REA 334496]. This work was supported by the Wellcome Trust [099149/Z/12/Z].
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