Sequential Protein Expression and Capsid Assembly In Cell: Towards the Study of Multi-Protein Viral Capsids Using Solid-State NMR Techniques.
Related Articles Sequential Protein Expression and Capsid Assembly In Cell: Towards the Study of Multi-Protein Viral Capsids Using Solid-State NMR Techniques.
Biochemistry. 2018 Feb 21;:
Authors: Alphonse S, Itin B, Khayat R, Ghose R
Abstract
While solid-state NMR (ssNMR) has emerged as a powerful technique to study viral capsids, current studies are limited to capsids formed from single proteins or single polyproteins. The ability to selectively label individual protein components within multi-protein 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 6 isotopically-labeled on only one of its four constituent protein components, the NTPase P4. Using P4-labeled 6 capsids and the sensitivity enhancement provided by dynamic nuclear polarization (DNP) we illustrate the utility of this method to enable ssNMR studies on complex viruses.
PMID: 29465229 [PubMed - as supplied by publisher]
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