Sequential Protein Expression and Capsid Assembly in Cell: Toward the Study of Multiprotein Viral Capsids Using Solid-State Nuclear Magnetic Resonance Techniques

Biochemistry
Sébastien AlphonseRanajeet Ghose

Abstract

While solid-state nuclear magnetic resonance (ssNMR) has emerged as a powerful technique for studying viral capsids, current studies are limited to capsids formed from single proteins or single polyproteins. The ability to selectively label individual protein components within multiprotein viral capsids and the resulting spectral simplification will facilitate the extension of ssNMR techniques to complex viruses. In vitro capsid assembly by combining individually purified, labeled, and unlabeled components in NMR quantities is not a viable option for most viruses. To overcome this barrier, we present a method that utilizes sequential protein expression and in cell assembly of component-specifically labeled viral capsids in amounts suitable for NMR studies. We apply this approach to purify capsids of bacteriophage ϕ6 isotopically labeled on only one of its four constituent protein components, the NTPase P4. Using P4-labeled ϕ6 capsids and the sensitivity enhancement provided by dynamic nuclear polarization, we illustrate the utility of this method to enable ssNMR studies of complex viruses.

References

Feb 7, 2007·Journal of the American Chemical Society·Amir GoldbourtAnn E McDermott
Jul 26, 2008·Proceedings of the National Academy of Sciences of the United States of America·Justin L LorieauAnn E McDermott
Jan 23, 2010·Journal of the American Chemical Society·Yun HanTatyana Polenova
Mar 31, 2011·Journal of Biomolecular NMR·Beomsoo HanDavid S Wishart
Oct 4, 2013·Journal of Virology·Xiaoyu SunMinna M Poranen
Aug 21, 2014·Biophysical Journal·Emeline Barbet-MassinGuido Pintacuda
Jan 15, 2015·Proceedings of the National Academy of Sciences of the United States of America·Omry MoragAmir Goldbourt
Apr 29, 2015·Progress in Nuclear Magnetic Resonance Spectroscopy·Caitlin M QuinnTatyana Polenova
Dec 29, 2015·The Journal of Physical Chemistry. B·Rupal GuptaTatyana E Polenova
Apr 25, 2017·The Journal of Physical Chemistry. B·Qing Zhe NiRobert G Griffin
May 4, 2017·Proceedings of the National Academy of Sciences of the United States of America·Ivan V SergeyevAnn E McDermott
Nov 28, 2017·Nature Communications·Mingzhang WangTatyana Polenova

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bacteriophage: Phage Therapy

Phage therapy uses bacterial viruses (bacteriophages) to treat bacterial infections and is widely being recognized as an alternative to antibiotics. Here is the latest research.