Probing dynamic conformations of the high-molecular-weight αB-crystallin heat shock protein ensemble by NMR spectroscopy.

Journal article

Solution- and solid-state nuclear magnetic resonance (NMR) spectroscopy are highly complementary techniques for studying supra-molecular structure. Here they are employed for investigating the molecular chaperone αB-crystallin, a polydisperse ensemble of between 10 and 40 identical subunits with an average molecular mass of approximately 600 kDa. An IxI motif in the C-terminal region of each of the subunits is thought to play a critical role in regulating the size distribution of oligomers and in controlling the kinetics of subunit exchange between them. Previously published solid-state NMR and X-ray results are consistent with a bound IxI conformation, while solution NMR studies provide strong support for a highly dynamic state. Here we demonstrate through FROSTY (freezing rotational diffusion of protein solutions at low temperature and high viscosity) MAS (magic angle spinning) NMR that both populations are present at low temperatures (<0 °C), while at higher temperatures only the mobile state is observed. Solution NMR relaxation dispersion experiments performed under physiologically relevant conditions establish that the motif interchanges between flexible (highly populated) and bound (sparsely populated) states. This work emphasizes the importance of using multiple methods in studies of supra-molecules, especially for highly dynamic ensembles where sample conditions can potentially affect the conformational properties observed.

Authors: Baldwin, A; Walsh, P; Hansen, D; Hilton, G; Benesch, J

• Publication status: Published
• Journal: Journal of the American Chemical Society
• Volume: 134
• Issue: 37
• Pages: 15343-15350
• Publication date: 2012-09-01
• DOI: 10.1021/ja307874r
• EISSN: 1520-5126
• ISSN: 0002-7863
• Language: English
• Keywords: Nuclear Magnetic Resonance, Biomolecular; Molecular Probes; Crystallins; Molecular Weight; Heat-Shock Proteins