The Francis Crick Institute
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CP-MAS and solution NMR studies of allosteric communication in CA-assemblies of HIV-1.

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journal contribution
posted on 2022-07-07, 10:33 authored by Giuseppe Nicastro, Massimo Lucci, Alain Oregioni, Geoff Kelly, Tom A Frenkiel, Ian A Taylor
Solution and solid-state NMR spectroscopy are highly complementary techniques for studying structure and dynamics in very high molecular weight systems. Here we have analysed the dynamics of HIV-1 capsid (CA) assemblies in presence of the cofactors IP6 and ATPγS and the host-factor CPSF6 using a combination of solution state and cross polarisation magic angle spinning (CP-MAS) solid-state NMR. In particular, dynamical effects on ns to µs and µs to ms timescales are observed revealing diverse motions in assembled CA. Using CP-MAS NMR, we exploited the sensitivity of the amide/Cα-Cβ backbone chemical shifts in DARR and NCA spectra to observe the plasticity of the HIV-1 CA tubular assemblies and also map the binding of cofactors and the dynamics of cofactor-CA complexes. In solution, we measured how the addition of host- and co-factors to CA -hexamers perturbed the chemical shifts and relaxation properties of CA-Ile and -Met methyl groups using transverse-relaxation-optimized NMR spectroscopy to exploit the sensitivity of methyl groups as probes in high-molecular weight proteins. These data show how dynamics of the CA protein assembly over a range of spatial and temporal scales play a critical role in CA function. Moreover, we show that binding of IP6, ATPγS and CPSF6 results in local chemical shift as well as dynamic changes for a significant, contiguous portion of CA, highlighting how allosteric pathways communicate ligand interactions between adjacent CA protomers.


Crick (Grant ID: 10178, Grant title: Taylor,I FC001178) Wellcome Trust (Grant ID: 108014/Z/15/Z, Grant title: WT 108014/Z/15/Z) Crick (Grant ID: 10029, Grant title: Frenkiel FC001029)