How to prepare membrane proteins for solid-state NMR: A case study on the alpha-helical integral membrane protein diacylglycerol kinase from E. coli.

@article{citeulike:1666146, abstract = {Several studies have demonstrated that it is viable to use microcrystalline preparations of water-soluble proteins as samples in solid-state NMR experiments [1-5]. Here, we investigate whether this approach holds any potential for studying water-insoluble systems, namely membrane proteins. For this case study, we have prepared proteoliposomes and small crystals of the alpha-helical membrane-protein diacylglycerol kinase (DGK). Preparations were characterised by 13C- and 15N-cross-polarization magic-angle spinning (CPMAS) NMR. It was found that crystalline samples produce better-resolved spectra than proteoliposomes. This makes them more suitable for structural NMR experiments. However, reconstitution is the method of choice for biophysical studies by solid-state NMR. In addition, we discuss the identification of lipids bound to membrane-protein crystals by 31P-MAS NMR.}, address = {Centre for Biomolecular Magnetic Resonance and Institut f\"{u}r Biophysikalische Chemie, J. W. Goethe Universit\"{a}t, Marie-Curie-Strasse 9, 60439 Frankfurt, Germany.}, author = {Lorch, M. and Fahem, S. and Kaiser, C. and Weber, I. and Mason, A. J. and Bowie, J. U. and Glaubitz, C. }, citeulike-article-id = {1666146}, doi = {10.1002/cbic.200500054}, issn = {1439-4227}, journal = {Chembiochem}, keywords = {dgk, membrane\_protein, ssnmr}, month = {September}, number = {9}, pages = {1693--1700}, posted-at = {2008-07-22 15:10:01}, priority = {2}, title = {How to prepare membrane proteins for solid-state NMR: A case study on the alpha-helical integral membrane protein diacylglycerol kinase from E. coli.}, url = {http://dx.doi.org/10.1002/cbic.200500054}, volume = {6}, year = {2005} }

TY - JOUR ID - citeulike:1666146 L3 - citeulike-article-id:1666146 TI - How to prepare membrane proteins for solid-state NMR: A case study on the alpha-helical integral membrane protein diacylglycerol kinase from E. coli. JF - Chembiochem VL - 6 IS - 9 SP - 1693 EP - 1700 AD - Centre for Biomolecular Magnetic Resonance and Institut für Biophysikalische Chemie, J. W. Goethe Universität, Marie-Curie-Strasse 9, 60439 Frankfurt, Germany. SN - 1439-4227 N2 - Several studies have demonstrated that it is viable to use microcrystalline preparations of water-soluble proteins as samples in solid-state NMR experiments [1-5]. Here, we investigate whether this approach holds any potential for studying water-insoluble systems, namely membrane proteins. For this case study, we have prepared proteoliposomes and small crystals of the alpha-helical membrane-protein diacylglycerol kinase (DGK). Preparations were characterised by 13C- and 15N-cross-polarization magic-angle spinning (CPMAS) NMR. It was found that crystalline samples produce better-resolved spectra than proteoliposomes. This makes them more suitable for structural NMR experiments. However, reconstitution is the method of choice for biophysical studies by solid-state NMR. In addition, we discuss the identification of lipids bound to membrane-protein crystals by 31P-MAS NMR. KW - dgk KW - membrane_protein KW - ssnmr AU - Lorch, M AU - Fahem, S AU - Kaiser, C AU - Weber, I AU - Mason, AJ AU - Bowie, JU AU - Glaubitz, C PY - 2005/09// UR - http://dx.doi.org/10.1002/cbic.200500054 ER -