Determining inter-domain structure and dynamics of a retroviral capsid protein in the presence of oligomerization: implication for structural transition in capsid assembly

摘要

Capsid (CA) proteins from all retroviruses, including HIV-1, are structurally homologous dual-domain helical proteins. They form a capsid lattice composed of unitary symmetric CA hexamers. X-ray crystallography has shown that within each hexamer a monomeric CA adopts a single conformation where most helices are parallel to the symmetry axis. In solution large differences in averaged NMR spin relaxation rates for the two domains were observed, suggesting they are dynamically independent. One relevant question for the capsid assembly remains, whether the inter-domain conformer within a hexamer unit needs to be induced or pre-exists within the conformational space of a monomeric CA. The latter seems more consistent with the relaxation data. However, possible CA protein oligomerization and the structure of each domain will affect relaxation measurements and data interpretation. Here using CA proteins from Equine Infectious Anemia Virus (EIAV) as an example, we demonstrate a linear-extrapolation approach to obtain backbone ¹⁵N spin relaxation time ratios T1/T2 for a monomeric EIAV-CA in the presence of oligomerization equilibrium. The inter-domain motion turns out to be limited. The large difference in the domain averaged <T1/T2> for a CA monomer is a consequence of the orthogonal distributions of helices in the two domains. The new monomeric inter-domain conformation in solution is significantly different from that in CA hexamer. Therefore if capsid assembly follows a nucleation-propagation process, the inter-domain conformational change might be a key step during the nucleation, as the configuration in hexagonal assembly is never formed by diffusion of its two domains in solution.