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dc.contributor.authorWang, Songlin
dc.contributor.authorParthasarathy, Sudhakar
dc.contributor.authorNishiyama, Yusuke
dc.contributor.authorEndo, Yuki
dc.contributor.authorNemoto, Takahiro
dc.contributor.authorYamauchi, Kazuo
dc.contributor.authorAsakura, Tetsuo
dc.contributor.authorTakeda, Mitsuhiro
dc.contributor.authorTerauchi, Tsutomu
dc.contributor.authorKainosho, Masatsune
dc.contributor.authorIshii, Yoshitaka
dc.date.accessioned2015-04-14T07:53:25Z
dc.date.available2015-04-14T07:53:25Z
dc.date.issued2015-04-09
dc.identifier.citationWang S, Parthasarathy S, Nishiyama Y, Endo Y, Nemoto T, et al. (2015) Nano-Mole Scale Side-Chain Signal Assignment by 1H-Detected Protein Solid-State NMR by Ultra-Fast Magic-Angle Spinning and Stereo-Array Isotope Labeling. PLoS ONE 10(4): e0122714. doi:10.1371/journal.pone.0122714
dc.identifier.issn1932-6203
dc.identifier.pmid25856081
dc.identifier.doi10.1371/journal.pone.0122714
dc.identifier.urihttp://hdl.handle.net/10754/550050
dc.description.abstractWe present a general approach in 1H-detected 13C solid-state NMR (SSNMR) for side-chain signal assignments of 10-50 nmol quantities of proteins using a combination of a high magnetic field, ultra-fast magic-angle spinning (MAS) at ~80 kHz, and stereo-array-isotope-labeled (SAIL) proteins [Kainosho M. et al., Nature 440, 52–57, 2006]. First, we demonstrate that 1H indirect detection improves the sensitivity and resolution of 13C SSNMR of SAIL proteins for side-chain assignments in the ultra-fast MAS condition. 1H-detected SSNMR was performed for micro-crystalline ubiquitin (~55 nmol or ~0.5mg) that was SAIL-labeled at seven isoleucine (Ile) residues. Sensitivity was dramatically improved by 1H-detected 2D 1H/13C SSNMR by factors of 5.4-9.7 and 2.1-5.0, respectively, over 13C-detected 2D 1H/13C SSNMR and 1D 13C CPMAS, demonstrating that 2D 1H-detected SSNMR offers not only additional resolution but also sensitivity advantage over 1D 13C detection for the first time. High 1H resolution for the SAIL-labeled side-chain residues offered reasonable resolution even in the 2D data. A 1H-detected 3D 13C/13C/1H experiment on SAIL-ubiquitin provided nearly complete 1H and 13C assignments for seven Ile residues only within ~2.5 h. The results demonstrate the feasibility of side-chain signal assignment in this approach for as little as 10 nmol of a protein sample within ~3 days. The approach is likely applicable to a variety of proteins of biological interest without any requirements of highly efficient protein expression systems.
dc.publisherPublic Library of Science (PLoS)
dc.relation.urlhttp://dx.plos.org/10.1371/journal.pone.0122714
dc.rightsThis is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
dc.titleNano-Mole Scale Side-Chain Signal Assignment by 1H-Detected Protein Solid-State NMR by Ultra-Fast Magic-Angle Spinning and Stereo-Array Isotope Labeling
dc.typeArticle
dc.contributor.departmentImaging and Characterization Core Lab
dc.identifier.journalPLoS ONE
dc.identifier.pmcidPMC4391754
dc.eprint.versionPublisher's Version/PDF
dc.contributor.institutionDepartment of Chemistry and University of Illinois at Chicago, Chicago, Illinois, United States of America
dc.contributor.institutionRIKEN CLST-JEOL collaboration center, RIKEN, Yokohama, Kanagawa, Japan
dc.contributor.institutionSchool of Science and Technology, Nazarbayev University, Astana, Kazakhstan
dc.contributor.institutionDepartment of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo, Japan
dc.contributor.institutionJEOL RESONANCE Inc., Akishima, Tokyo, Japan
dc.contributor.institutionStructural Biology Research Center, Graduate School of Science, Furocho, Chikusa-ku, Nagoya University, Nagoya, Japan 464–8601
dc.contributor.institutionSAIL Technologies Co., Inc., Tsurumi-ku, Yokohama, Kanagawa, Japan
dc.contributor.institutionCenter for Priority Areas, Tokyo Metropolitan University, Tokyo, Japan
dc.contributor.institutionCenter for Structural Biology, University of Illinois at Chicago, Chicago, Illinois, United States of America
kaust.personYamauchi, Kazuo
refterms.dateFOA2018-06-13T16:48:50Z


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