caa a22 4500 44532516X CHVBK 20180317142718.0 cr unu---uuuuu 170323e20111101xx s 000 0 eng 10.1007/s10858-011-9565-6 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9565-6 High-resolution membrane protein structure by joint calculations with solid-state NMR and X-ray experimental data [Elektronische Daten] [Ming Tang, Lindsay Sperling, Deborah Berthold, Charles Schwieters, Anna Nesbitt, Andrew Nieuwkoop, Robert Gennis, Chad Rienstra] X-ray diffraction and nuclear magnetic resonance spectroscopy (NMR) are the staple methods for revealing atomic structures of proteins. Since crystals of biomolecular assemblies and membrane proteins often diffract weakly and such large systems encroach upon the molecular tumbling limit of solution NMR, new methods are essential to extend structures of such systems to high resolution. Here we present a method that incorporates solid-state NMR restraints alongside of X-ray reflections to the conventional model building and refinement steps of structure calculations. Using the 3.7Å crystal structure of the integral membrane protein complex DsbB-DsbA as a test case yielded a significantly improved backbone precision of 0.92Å in the transmembrane region, a 58% enhancement from using X-ray reflections alone. Furthermore, addition of solid-state NMR restraints greatly improved the overall quality of the structure by promoting 22% of DsbB transmembrane residues into the most favored regions of Ramachandran space in comparison to the crystal structure. This method is widely applicable to any protein system where X-ray data are available, and is particularly useful for the study of weakly diffracting crystals. Springer Science+Business Media B.V., 2011 Membrane protein nationallicence Solid-state NMR nationallicence X-ray reflections nationallicence High resolution nationallicence Joint calculation nationallicence Tang Ming Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, 61801, Urbana, IL, USA aut Sperling Lindsay Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, 61801, Urbana, IL, USA aut Berthold Deborah Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, 61801, Urbana, IL, USA aut Schwieters Charles Division of Computational Bioscience, Center for Information Technology, National Institutes of Health, 20892, Bethesda, MD, USA aut Nesbitt Anna Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, 61801, Urbana, IL, USA aut Nieuwkoop Andrew Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, 61801, Urbana, IL, USA aut Gennis Robert Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, 61801, Urbana, IL, USA aut Rienstra Chad Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, 61801, Urbana, IL, USA aut Journal of Biomolecular NMR Springer Netherlands 51/3(2011-11-01), 227-233 0925-2738 51:3<227 2011 51 10858 https://doi.org/10.1007/s10858-011-9565-6 text/html Onlinezugriff via DOI 1 brief-communication jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9565-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Tang Ming Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, 61801, Urbana, IL, USA aut NATIONALLICENCE 700 1- Sperling Lindsay Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, 61801, Urbana, IL, USA aut NATIONALLICENCE 700 1- Berthold Deborah Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, 61801, Urbana, IL, USA aut NATIONALLICENCE 700 1- Schwieters Charles Division of Computational Bioscience, Center for Information Technology, National Institutes of Health, 20892, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Nesbitt Anna Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, 61801, Urbana, IL, USA aut NATIONALLICENCE 700 1- Nieuwkoop Andrew Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, 61801, Urbana, IL, USA aut NATIONALLICENCE 700 1- Gennis Robert Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, 61801, Urbana, IL, USA aut NATIONALLICENCE 700 1- Rienstra Chad Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, 61801, Urbana, IL, USA aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 51/3(2011-11-01), 227-233 0925-2738 51:3<227 2011 51 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer