An evaluation of detergents for NMR structural studies of membrane proteins

Ray D. Krueger-Koplin, Paul L. Sorgen, Suzanne T. Krueger-Koplin, Iván O. Rivera-Torres, Sean M. Cahill, David B. Hicks, Leo Grinius, Terry A. Krulwich, Mark E. Girvin

Research output: Contribution to journalArticlepeer-review

153 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)43-57
Number of pages15
JournalJournal of Biomolecular NMR
Volume28
Issue number1
DOIs
StatePublished - Jan 2004
Externally publishedYes

Keywords

  • Detergent
  • Lysolipids
  • Membrane
  • Protein
  • Structure

ASJC Scopus subject areas

  • Biochemistry
  • Spectroscopy

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