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Preparation, characterization, and NMR spectroscopy of encapsulated proteins dissolved in low viscosity fluids

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Abstract

Encapsulating a protein in a reverse micelle and dissolving it in a low-viscosity solvent can lower the rotational correlation time of a protein and thereby provides a novel strategy for studying proteins in a variety of contexts. The preparation of the sample is a key element in this approach and is guided by a number of competing parameters. Here we examine the applicability of several strategies for the preparation and characterization of encapsulated proteins dissolved in low viscosity fluids that are suitable for high performance NMR spectroscopy. Ubiquitin is used as a model system to explore various issues such as the homogeneity of the encapsulation, characterization of the hydrodynamic performance of reverse micelles containing protein molecules, and the effective pH of the water environment of the reverse micelle.

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Authors and Affiliations

  1. The Johnson Research Foundation and Department of Biochemistry & Biophysics, University of Pennsylvania, Philadelphia, Pennsylvania, 19104, U.S.A

    Charles R. Babu, Peter F. Flynn & A. Joshua Wand

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  1. Charles R. Babu
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  2. Peter F. Flynn
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  3. A. Joshua Wand
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Correspondence to A. Joshua Wand.

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Babu, C.R., Flynn, P.F. & Wand, A.J. Preparation, characterization, and NMR spectroscopy of encapsulated proteins dissolved in low viscosity fluids. J Biomol NMR 25, 313–323 (2003). https://doi.org/10.1023/A:1023037823421

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