Translational Diffusion of Macromolecular Assemblies Measured Using Transverse-Relaxation-Optimized Pulsed Field Gradient NMR

In structural biology, pulsed field gradient (PFG) NMR spectroscopy for the characterization of size and hydrodynamic parameters of macromolecular solutes has the advantage over other techniques that the measurements can be recorded with identical solution conditions as used for NMR structure determ...

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Bibliographic Details
Published in:Journal of the American Chemical Society 2011-10, Vol.133 (41), p.16354-16357
Main Authors: Horst, Reto, Horwich, Arthur L, Wüthrich, Kurt
Format: Article
Language:English
Subjects:
Bacterial Outer Membrane Proteins - chemistry
Chaperonin 60 - chemistry
Detergents - chemistry
Diffusion
Escherichia coli - chemistry
Escherichia coli Proteins - chemistry
Hydrolases - chemistry
Macromolecular Substances - chemistry
Magnetic Resonance Spectroscopy
Thermus thermophilus - chemistry
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Summary:In structural biology, pulsed field gradient (PFG) NMR spectroscopy for the characterization of size and hydrodynamic parameters of macromolecular solutes has the advantage over other techniques that the measurements can be recorded with identical solution conditions as used for NMR structure determination or for crystallization trials. This paper describes two transverse-relaxation-optimized (TRO) 15N-filtered PFG stimulated-echo (STE) experiments for studies of macromolecular translational diffusion in solution, 1H-TRO-STE and 15N-TRO-STE, which include CRINEPT and TROSY elements. Measurements with mixed micelles of the Escherichia coli outer membrane protein X (OmpX) and the detergent Fos-10 were used for a systematic comparison of 1H-TRO-STE and 15N-TRO-STE with conventional 15N-filtered STE experimental schemes. The results provide an extended platform for evaluating the NMR experiments available for diffusion measurements in structural biology projects involving molecular particles with different size ranges. An initial application of the 15N-TRO-STE experiment with very long diffusion delays showed that the tedradecamer structure of the 800 kDa Thermus thermophilus chaperonin GroEL is preserved in aqueous solution over the temperature range 25–60 °C.
ISSN:0002-7863
1520-5126
DOI:10.1021/ja206531c