FLYA

Fully automated structure determination of proteins in solution

Fully automated structure determination of proteins in solution (FLYA) yields, without human intervention, three-dimensional protein structures starting from a set of multidimensional NMR spectra. The core FLYA algorithm determines chemical shift assignments for all ¹H, ¹³C, and ¹⁵N nuclei, the assignment of NOESY cross-peaks, the generation of distance restraints, and the calculation of the three-dimensional structure by torsion angle dynamics. The resulting, preliminary structure serves as additional input to the second stage of the procedure, in which a new ensemble of chemical shift assignments and a refined structure are calculated. The purely computational FLYA method is suitable for substituting all manual spectra analysis and thus overcomes a main efficiency limitation of the NMR method for protein structure determination.

Availability

• FLYA calculations are carried out with the program CYANA. FLYA is not available as a separate program.

References

FLYA algorithm:

• Schmidt, E. & Güntert, P. A new algorithm for reliable and general NMR resonance assignment. J. Am. Chem. Soc. 134, 12817–12829 (2012)

FLYA applications:

• Schmidt, E., Gath, J., Habenstein, B., Ravotti, F., Székely, K., Huber, M., Buchner, L., Böckmann, A., Meier, B. H. & Güntert, P. Automated solid-state NMR resonance assignment of protein microcrystals and amyloids. J. Biomol. NMR 56, 243–254 (2013)

• Schmidt, E. & Güntert, P. Reliability of exclusively NOESY-based automated resonance assignment and structure determination of proteins. J. Biomol. NMR 57, 193-204 (2013)

• Aeschbacher, T., Schmidt, E., Blatter, M., Maris, C., Duss, O., Allain, F. H.-T., Güntert, P. & Schubert, M. Automated and assisted RNA resonance assignment using NMR chemical shift statistics. Nucl. Acids Res. 41, e172 (2013)

• Krähenbühl, B., El Bakkali, I., Schmidt, E., Güntert, P. & Wider, G. Automated NMR resonance assignment strategy for RNA via the phosphodiester backbone based on high-dimensional through-bond APSY experiments. J. Biomol. NMR 59, 87-93 (2014)

• Schmidt, E., Ikeya, T., Takeda, M., Löhr, F., Buchner, L., Ito, Y., Kainosho, M. & Güntert, P. Automated resonance assignment of the 21 kDa stereo-array isotope labeled thioldisulfide oxidoreductase DsbA. J. Magn. Reson. 249, 88–93 (2014)

• Schmidt, E. & Güntert, P. Automated structure determination from NMR spectra. Meth. Mol. Biol. 1261, 303–329 (2015)

Combined automated NOESY cross peak assignment and structure calculation:

• Güntert, P. & Buchner, L. Combined automated NOE assignment and structure calculation with CYANA. J. Biomol. NMR 62, 453-471 (2015)

Evaluation of combined automated NOESY cross peak assignment and structure calculation

• Buchner, L. & Güntert, P. Systematic evaluation of combined automated NOE assignment and structure calculation with CYANA. J. Biomol. NMR 62, 81–95 (2015)