APSY-NMR for protein backbone assignment in high-throughput structural biology
- Journal Article
Rights / licenseIn Copyright - Non-Commercial Use Permitted
A standard set of three APSY-NMR experiments has been used in daily practice to obtain polypeptide backbone NMR assignments in globular proteins with sizes up to about 150 residues, which had been identified as targets for structure determination by the Joint Center for Structural Genomics (JCSG) under the auspices of the Protein Structure Initiative (PSI). In a representative sample of 30 proteins, initial fully automated data analysis with the software UNIO-MATCH-2014 yielded complete or partial assignments for over 90 % of the residues. For most proteins the APSY data acquisition was completed in less than 30 h. The results of the automated procedure provided a basis for efficient interactive validation and extension to near-completion of the assignments by reference to the same 3D heteronuclear-resolved [1H,1H]-NOESY spectra that were subsequently used for the collection of conformational constraints. High-quality structures were obtained for all 30 proteins, using the J-UNIO protocol, which includes extensive automation of NMR structure determination. Show more
Journal / seriesJournal of Biomolecular NMR
Pages / Article No.
SubjectAutomated data analysis; Automated projection spectroscopy; J-UNIO protocol; Protein structure determination; UNIO software
NotesIt was possible to publish this article open access thanks to a Swiss National Licence with the publisher.
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