Progress in FY2012 was in the following areas: (1) HIV-1 CAPSID ASSEMBLIES. A new postdoctoral fellow, Marvin Bayro, began working on solid state NMR studies of HIV-1 capsid (CA) assemblies at the beginning of FY12. He has acquired a large set of 2D and 3D solid state NMR spectra of tubular CA assemblies, with both uniformly 15N,13C-labeled CA and with partially labeled CA, grown on selectively labeled glycerol media. We are in the process of analyzing these spectra to determine site-specific resonance assignments, which will tell us which segments of CA are structurally ordered and immobilized in CA assemblies, and which residues are involved in conformational changes upon formation of these assemblies. Dr. Bayro has also developed a protocol for preparation of segmentally labeled CA (labeled either in N-terminal or C-terminal sections), which will provide great simplification of the solid state NMR data. (2) MEMBRANE-ASSOCIATED HIV-1 MATRIX PROTEIN. We have performed initial solid state NMR measurements to assess the feasibility of studies to investigate intermolecular interactions of membrane-bound HIV-1 MA proteins. These experiments will be pursued in FY13. (3) HEPATITIS B VIRUS CAPSID STRUCTURE. In collaboration with Dr. Norman Watts, we have acquired 2D and 3D solid state NMR spectra of uniformly 15N,13C-labeled HBV capsid protein, assembled into capsid particles with icosahedral symmetry. Analysis of the data is in progress. We expect to obtain new information about site-specific dynamics and site-specific structural variations that underlie quasi-equivalence in T=3 and T=4 capsid particles.

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Gupta, Sebanti; Tycko, Robert (2018) Segmental isotopic labeling of HIV-1 capsid protein assemblies for solid state NMR. J Biomol NMR 70:103-114
Bayro, Marvin J; Ganser-Pornillos, Barbie K; Zadrozny, Kaneil K et al. (2016) Helical Conformation in the CA-SP1 Junction of the Immature HIV-1 Lattice Determined from Solid-State NMR of Virus-like Particles. J Am Chem Soc 138:12029-32
Bayro, Marvin J; Tycko, Robert (2016) Structure of the Dimerization Interface in the Mature HIV-1 Capsid Protein Lattice from Solid State NMR of Tubular Assemblies. J Am Chem Soc 138:8538-46
Lu, Jun-Xia; Bayro, Marvin J; Tycko, Robert (2016) Major Variations in HIV-1 Capsid Assembly Morphologies Involve Minor Variations in Molecular Structures of Structurally Ordered Protein Segments. J Biol Chem 291:13098-112
Bayro, Marvin J; Chen, Bo; Yau, Wai-Ming et al. (2014) Site-specific structural variations accompanying tubular assembly of the HIV-1 capsid protein. J Mol Biol 426:1109-27
Chen, Bo; Tycko, Robert (2011) Simulated self-assembly of the HIV-1 capsid: protein shape and native contacts are sufficient for two-dimensional lattice formation. Biophys J 100:3035-44
Lu, Jun-Xia; Sharpe, Simon; Ghirlando, Rodolfo et al. (2010) Oligomerization state and supramolecular structure of the HIV-1 Vpu protein transmembrane segment in phospholipid bilayers. Protein Sci :
Chen, Bo; Tycko, Robert (2010) Structural and dynamical characterization of tubular HIV-1 capsid protein assemblies by solid state nuclear magnetic resonance and electron microscopy. Protein Sci 19:716-30