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 journalArticle

147 Citations (Scopus)

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

Fingerprint

Detergents
Membrane Proteins
Nuclear magnetic resonance
Membranes
Proteins
Barreling
Micelles
Multiple Drug Resistance
Dimers
Labeling
Escherichia coli
Spectrum Analysis
Adenosine Triphosphate
Pharmaceutical Preparations
Experiments

Keywords

  • Detergent
  • Lysolipids
  • Membrane
  • Protein
  • Structure

ASJC Scopus subject areas

  • Spectroscopy
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Krueger-Koplin, R. D., Sorgen, P. L., Krueger-Koplin, S. T., Rivera-Torres, I. O., Cahill, S. M., Hicks, D. B., ... Girvin, M. E. (2004). An evaluation of detergents for NMR structural studies of membrane proteins. Journal of Biomolecular NMR, 28(1), 43-57. https://doi.org/10.1023/B:JNMR.0000012875.80898.8f

An evaluation of detergents for NMR structural studies of membrane proteins. / Krueger-Koplin, Ray D.; Sorgen, Paul L.; Krueger-Koplin, Suzanne T.; Rivera-Torres, Iván O.; Cahill, Sean M.; Hicks, David B.; Grinius, Leo; Krulwich, Terry A.; Girvin, Mark E.

In: Journal of Biomolecular NMR, Vol. 28, No. 1, 01.2004, p. 43-57.

Research output: Contribution to journalArticle

Krueger-Koplin, RD, Sorgen, PL, Krueger-Koplin, ST, Rivera-Torres, IO, Cahill, SM, Hicks, DB, Grinius, L, Krulwich, TA & Girvin, ME 2004, 'An evaluation of detergents for NMR structural studies of membrane proteins', Journal of Biomolecular NMR, vol. 28, no. 1, pp. 43-57. https://doi.org/10.1023/B:JNMR.0000012875.80898.8f
Krueger-Koplin RD, Sorgen PL, Krueger-Koplin ST, Rivera-Torres IO, Cahill SM, Hicks DB et al. An evaluation of detergents for NMR structural studies of membrane proteins. Journal of Biomolecular NMR. 2004 Jan;28(1):43-57. https://doi.org/10.1023/B:JNMR.0000012875.80898.8f
Krueger-Koplin, Ray D. ; Sorgen, Paul L. ; Krueger-Koplin, Suzanne T. ; Rivera-Torres, Iván O. ; Cahill, Sean M. ; Hicks, David B. ; Grinius, Leo ; Krulwich, Terry A. ; Girvin, Mark E. / An evaluation of detergents for NMR structural studies of membrane proteins. In: Journal of Biomolecular NMR. 2004 ; Vol. 28, No. 1. pp. 43-57.
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