Columbus L, Lipfert J, Jambunathan K, Fox DA, Sim AYL, Doniach S, and Lesley SA.
J. Am. Chem. Soc., 2009, 131 (21), pp 7320–7326
One major obstacle to membrane protein structure determination is the selection of a detergent micelle that mimics the native lipid bilayer. Currently, detergents are selected by exhaustive screening because the effects of protein−detergent interactions on protein structure are poorly understood. In this study, the structure and dynamics of an integral membrane protein in different detergents is investigated by nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR) spectroscopy and small-angle X-ray scattering (SAXS). The results suggest that matching of the micelle dimensions to the protein’s hydrophobic surface avoids exchange processes that reduce the completeness of the NMR observations. Based on these dimensions, several mixed micelles were designed that improved the completeness of NMR observations. These findings provide a basis for the rational design of mixed micelles that may advance membrane protein structure determination by NMR.

Columbus L, Nakamoto, RK, Cafiso, DS. Chapter in Wiley Encyclopedia of Chemical Biology (2008)

McCleverty C*, Columbus L*, Kreusch A, Lesley SA. Structure and ligand binding of the soluble domain of a Thermotoga maritima membrane protein of unknown function TM1634. Protein Science in press (2008)

Lipfert J, Columbus L, Chu V, Lesley SA, Doniach S. Size and shape of detergent micelles determined by small-angle X-ray scattering. Journal of Physical Chemistry B 111: 12427-12438 (2007)

Lipfert J, Columbus L, Chu V, Doniach S. Analysis of small-angle X-ray scattering data of protein-detergent complexes with singular value decomposition. Journal of Applied Crystallography 40: s235-s239 (2007)

Columbus L, Lipfert J, Klock H, Millet I, Doniach S, Lesley SA. Expression, purification, and characterization of Thermotoga maritimamembrane proteins for structure determination. Protein Science 15: 961-975 (2006)

Columbus L, Peti W, Herrmann T, Etezady T, Wüthrich K. NMR structure determination of the conserved hypothetical protein TM1816 from Thermotoga maritima . Proteins: Structure, Function and Bioinformatics 60: 552-557 (2005)

Liang ZC, Lou Y, Freed JH, Columbus L, Hubbell WL. A multifrequency electron spin resonance study of T4 lysozyme dynamics using the slowly relaxing local structure model Journal of Physical Chemistry B 108: 17649-17659 (2004)

Columbus L, Hubbell WL. Mapping backbone dynamics in solution with site-directed spin labeling: GCN4-58 bZip free and bound to DNA. Biochemistry 43: 7273-7287 (2004)

Columbus L, Hubbell WL. A new spin on protein dynamics. Trends in Biochemical Sciences, 27: 288-295 (2002)

Columbus L, Kalai T, Jeko J, Hideg K, Hubbell WL. Molecular motion of spin labeled side chains in α-helices: Analysis by variation of side chain structure. Biochemistry 40: 3828-3846 (2001)

Gaponenko V, Howarth JW, Columbus L, Gasmi-Seabrook G, Yuan J, Hubbell WL, Rosevear PR. Protein global fold determination using site-directed spin and isotope labeling. Protein Science 9: 302-309 (2000)

Gross A, Columbus L, Hideg K, Altenbach C, Hubbell WL. Structure of the KcsA potassium channel from Streptomyces lividans: A site-directed spin labeling study of the second transmembrane segment. Biochemistry 38: 10324-10335 (1999)