AI|ffinity NMR
4D-GRAPHS enhances protein structure determination by using 4D NMR, resolving spectral crowding and reducing experimental time. It efficiently determines mutant protein structures with fewer spectra, lowering costs.
Protein structure determination by NMR is a complex, expensive, and time-consuming process. The traditional approach requires recording 12 to 16 3D NMR spectra, which becomes impractical for proteins larger than 20 kDa due to spectral crowding.
In this project, we upgraded, applied, and benchmarked new software based on 4D NMR spectra named 4D-GRAPHS. The extra dimension helps resolve crowded peaks and reduces experimental time by condensing the same information into fewer spectra.
Additionally, we addressed the challenge of mutant protein structure determination. Since mutant proteins share most of their chemical shifts with wild-type proteins with differences limited in the proximity of the mutations, 4D-GRAPHS can determine their structures with even fewer spectra, further improving efficiency and reducing costs. 4D-GRAPHS was tested on carbonic anhydrase 2 (CA2) and its mutant CA29, both weighting 29 kDa, as well as on wilde type HSP90 and it's mutants L129I and L103I, all weighting 30 kDa.
The project is funded by the OP-TAK Proof of Concept – Activity a) – Call I. (Project Number CZ.01.01.01/08/22_001/0000343) and contributes to advancing the fields of structural biology and drug discovery.
