An evaluation of detergents for NMR structural studies of membrane proteins Export

@article{citeulike:1557179, 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 a and {\ss} 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 a-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 {\ss} 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.}, author = {Krueger-Koplin, Ray D. and Sorgen, Paul L. and Krueger-Koplin, Suzanne T. and Rivera-Torres, Iv\'{a}n O. and Cahill, Sean M. and Hicks, David B. and Grinius, Leo and Krulwich, Terry A. and Girvin, Mark E.}, citeulike-article-id = {1557179}, citeulike-linkout-0 = {http://dx.doi.org/10.1023/B:JNMR.0000012875.80898.8f}, citeulike-linkout-1 = {http://www.ingentaconnect.com/content/klu/jnmr/2004/00000028/00000001/05147894}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/14739638}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=14739638}, day = {1}, doi = {10.1023/B:JNMR.0000012875.80898.8f}, journal = {Journal of Biomolecular NMR}, keywords = {detergent, detergents, lppg, membrane, nmr, protein, proteins, structure}, month = {January}, number = {1}, pages = {43--57}, posted-at = {2008-09-24 06:17:05}, priority = {2}, title = {An evaluation of detergents for NMR structural studies of membrane proteins}, url = {http://dx.doi.org/10.1023/B:JNMR.0000012875.80898.8f}, volume = {28}, year = {2004} }

TY - JOUR ID - citeulike:1557179 L3 - citeulike-article-id:1557179 N2 - 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 a and ß 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 a-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 ß 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. IS - 1 JF - Journal of Biomolecular NMR EP - 57 TI - An evaluation of detergents for NMR structural studies of membrane proteins VL - 28 SP - 43 KW - detergent KW - detergents KW - lppg KW - membrane KW - nmr KW - protein KW - proteins KW - structure AU - Krueger-Koplin, Ray AU - Sorgen, Paul AU - Krueger-Koplin, Suzanne AU - Rivera-Torres, Iván AU - Cahill, Sean AU - Hicks, David AU - Grinius, Leo AU - Krulwich, Terry AU - Girvin, Mark PY - 2004/01/01/ UR - http://dx.doi.org/10.1023/B:JNMR.0000012875.80898.8f ER -