TY - JOUR
T1 - The J-UNIO protocol for automated protein structure determination by NMR in solution
AU - Serrano, Pedro
AU - Pedrini, Bill
AU - Mohanty, Biswaranjan
AU - Geralt, Michael
AU - Herrmann, Torsten
AU - Wüthrich, Kurt
PY - 2012/8
Y1 - 2012/8
N2 - The J-UNIO (JCSG protocol using the software UNIO) procedure for automated protein structure determination by NMR in solution is introduced. In the present implementation, J-UNIO makes use of APSY-NMR spectroscopy, 3D heteronuclear-resolved [1H,1H]-NOESY experiments, and the software UNIO. Applications with proteins from the JCSG target list with sizes up to 150 residues showed that the procedure is highly robust and efficient. In all instances the correct polypeptide fold was obtained in the first round of automated data analysis and structure calculation. After interactive validation of the data obtained from the automated routine, the quality of the final structures was comparable to results from interactive structure determination. Special advantages are that the NMR data have been recorded with 6-10 days of instrument time per protein, that there is only a single step of chemical shift adjustments to relate the backbone signals in the APSY-NMR spectra with the corresponding backbone signals in the NOESY spectra, and that the NOE-based amino acid side chain chemical shift assignments are automatically focused on those residues that are heavily weighted in the structure calculation. The individual working steps of J-UNIO are illustrated with the structure determination of the protein YP-926445.1 from Shewanella amazonensis, and the results obtained with 17 JCSG targets are critically evaluated.
AB - The J-UNIO (JCSG protocol using the software UNIO) procedure for automated protein structure determination by NMR in solution is introduced. In the present implementation, J-UNIO makes use of APSY-NMR spectroscopy, 3D heteronuclear-resolved [1H,1H]-NOESY experiments, and the software UNIO. Applications with proteins from the JCSG target list with sizes up to 150 residues showed that the procedure is highly robust and efficient. In all instances the correct polypeptide fold was obtained in the first round of automated data analysis and structure calculation. After interactive validation of the data obtained from the automated routine, the quality of the final structures was comparable to results from interactive structure determination. Special advantages are that the NMR data have been recorded with 6-10 days of instrument time per protein, that there is only a single step of chemical shift adjustments to relate the backbone signals in the APSY-NMR spectra with the corresponding backbone signals in the NOESY spectra, and that the NOE-based amino acid side chain chemical shift assignments are automatically focused on those residues that are heavily weighted in the structure calculation. The individual working steps of J-UNIO are illustrated with the structure determination of the protein YP-926445.1 from Shewanella amazonensis, and the results obtained with 17 JCSG targets are critically evaluated.
KW - H-H-NOE
KW - APSY-NMR
KW - Automation
KW - JCSG targets
KW - Joint Center for Structural Genomics (JCSG)
KW - Protein structure initiative (PSI)
KW - UNIO software
UR - http://www.scopus.com/inward/record.url?scp=84865127233&partnerID=8YFLogxK
U2 - 10.1007/s10858-012-9645-2
DO - 10.1007/s10858-012-9645-2
M3 - Article
C2 - 22752932
AN - SCOPUS:84865127233
SN - 0925-2738
VL - 53
SP - 341
EP - 354
JO - Journal of Biomolecular NMR
JF - Journal of Biomolecular NMR
IS - 4
ER -