NMR assignment of the nonstructural protein nsp3(1066–1181) from SARS-CoV
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Sequence-specific NMR assignments of the globular core comprising the residues 1066–1181 within the non-structural protein nsp3e from the SARS coronavirus have been obtained using triple-resonance NMR experiments with the uniformly [13C, 15N]-labeled protein. The backbone and side chain assignments are nearly complete, providing the basis for the ongoing NMR structure determination. A preliminary identification of regular secondary structures has been derived from the 13C chemical shifts.
KeywordsSevere acute respiratory syndrome SARS coronavirus Nonstructural protein NMR structure determination
We thank Jeremiah Joseph, Vanitha Subramanian, Benjamin W. Neuman, Michael J. Buchmeier, Raymond C. Stevens, and Peter Kuhn of the Consortium for Functional and Structural Proteomics of the SARS-CoV for providing us with samples of nsp3(1066–1226) for the initial NMR screening. This study was supported by the NIAID/NIH contract #HHSN266200400058C “Functional and Structural Proteomics of the SARS-CoV” to P. Kuhn and M. J. Buchmeier, and by the Joint Center for Structural Genomics through the NIH/NIGMS Grant #U54-GM074898. Additional support was obtained for P. S., M. A. J. and B. P. through fellowships from the Spanish Ministry of Science and Education, the Canadian Institutes of Health Research, and the Swiss National Science Foundation (fellowship PA00A-109047/1), respectively, and by the Skaggs Institute for Chemical Biology. Kurt Wüthrich is the Cecil H. and Ida M. Green Professor of Structural Biology at TSRI.
- Metzler WJ, Constantine KL, Friedrichs MS, Bell AJ, Ernst EG, Lavoie TB, et al (1993) Characterization of the three-dimensional solution structure of human profilin: 1H, 13C, and 15N NMR assignments and global folding pattern. Biochemistry 32:13818–13829. doi: 10.1021/bi00213a010 CrossRefGoogle Scholar
- Pastore A, Saudek V (1990) The relationship between chemical shift and secondary structure in proteins. J Magn Reson 90:165–176Google Scholar
- Saikatendu KS, Joseph JS, Subramanian V, Clayton T, Griffith M, Moy K, et al (2005) Structural basis of severe acute respiratory syndrome coronavirus ADP-ribose-1′′-phosphate dephosphorylation by a conserved domain of nsp3. Structure 13:1665–1675. doi: 10.1016/j.str.2005.07.022 CrossRefGoogle Scholar