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An evaluation of detergents for NMR structural studies of membrane proteins

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Abstract

Structural information on membrane proteins lags far behind that on soluble proteins, in large part due to difficulties producing homogeneous, stable, structurally relevant samples in a membrane-like environment. In this study 25 membrane mimetics were screened using 2D 1H-15N heteronuclear single quantum correlation NMR experiments to establish sample homogeneity and predict fitness for structure determination. A single detergent, 1-palmitoyl-2-hydroxy-sn-glycero-3-[phospho-RAC-(1-glycerol)] (LPPG), yielded high quality NMR spectra with sample lifetimes greater than one month for the five proteins tested – R. sphaeroides LH1 α and β subunits, E. coli and B. pseudofirmus OF4 ATP synthase c subunits, and S. aureus small multidrug resistance transporter – with 1, 2, or 4 membrane spanning α-helices, respectively. Site-specific spin labeling established interhelical distances in the drug transporter and genetically fused dimers of c subunits in LPPG consistent with in vivo distances. Optical spectroscopy showed that LH1 β subunits form native-like complexes with bacteriochlorphyll a in LPPG. All the protein/micelle complexes were estimated to exceed 100 kDaltons by translational diffusion measurements. However, analysis of 15N transverse, longitudinal and 15N{1H} nuclear Overhauser effect relaxation measurements yielded overall rotational correlation times of 8 to 12 nsec, similar to a 15–20 kDalton protein tumbling isotropically in solution, and consistent with the high quality NMR data observed.

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Authors and Affiliations

  1. Biochemistry Department, Albert Einstein College of Medicine, 1300 Morris Park Ave, Bronx, NY, 10461, U.S.A.

    Ray D. Krueger-Koplin, Paul L. Sorgen, Suzanne T. Krueger-Koplin, Iván O. Rivera-Torres, Sean M. Cahill & Mark E. Girvin

  2. Department of Pharmacology and Biological Chemistry, Mt, Sinai School of Medicine, One Gustave L. Levy Place, Box 1603, New York, NY, 10029, U.S.A.

    David B. Hicks & Terry A. Krulwich

  3. Cincinnati State Technical College, 3520 Central Parkway, Cincinnati, Ohio, 45223, U.S.A.

    Leo Grinius

Authors
  1. Ray D. Krueger-Koplin
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  2. Paul L. Sorgen
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  3. Suzanne T. Krueger-Koplin
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  4. Iván O. Rivera-Torres
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  5. Sean M. Cahill
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  6. David B. Hicks
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  7. Leo Grinius
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  8. Terry A. Krulwich
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  9. Mark E. Girvin
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Krueger-Koplin, R.D., Sorgen, P.L., Krueger-Koplin, S.T. et al. An evaluation of detergents for NMR structural studies of membrane proteins. J Biomol NMR 28, 43–57 (2004). https://doi.org/10.1023/B:JNMR.0000012875.80898.8f

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  • DOI: https://doi.org/10.1023/B:JNMR.0000012875.80898.8f

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