APSY-NMR for protein backbone assignment in high-throughput structural biology
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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.
KeywordsAutomated projection spectroscopy Automated data analysis UNIO software J-UNIO protocol Protein structure determination
- Jaudzems K, Pedrini B, Geralt M, Serrano P, Wüthrich K (2014) J-UNIO protocol used for NMR structure determination of the 206-residue protein NP_346487.1 from Streptococcus pneumoniae TIGR4. J Biomol NMR. doi:10.1007/s10858-014-9886-3
- Keller R (2004) Computer-aided resonance assignment. Cantina. http://cara.nmr.ch/
- Mohanty B, Serrano P, Geralt M, Wüthrich K (2014) NMR structure determination of the protein NP_344798.1 as the first representative of Pfam PF06042. J Biomol NMR. doi:10.1007/s10858-014-9878-3
- Wüthrich K (1986) NMR of proteins and nucleic acids. Wiley, New YorkGoogle Scholar