Transforming biomolecular NMR to stay at the forefront of Structural Biology

Konstantinos Tripsianes

CEITEC — Central European Institute of Technology, Masaryk University, Kamenice 5, Brno 62500, Czech Republic, kostas.tripsianes@ceitec.muni.cz

 

The automation of NMR structure determination remains a significant bottleneck towards increasing the throughput and accessibility of NMR as a structural biology tool to study proteins. The chief barrier currently is that obtaining NMR assignments at sufficient levels of completeness to accurately define the structures by conventional methods requires a significant amount of spectrometer time (several weeks), and effort by a trained expert (up to several months). We have recently addressed both bottlenecks by presenting a complete pipeline for NMR structure determination using a minimal set of NMR spectra. Key to our approach was the development of 4D-CHAINS algorithm that enables fully automated assignments of NMR chemical shifts, at high levels of completeness and with a minimum error rate, from only two complementary spectra. In combination with autoNOE-Rosetta, 4D-CHAINS provides a robust approach leveraging a highly automated process to obtain reliable structures in a matter of days. Besides illustrating the merits of our pipeline for timely NMR structural studies, novel concepts in automation will be discussed aiming to harness the powerful advantages of the next-generation NMR spectrometers with magnetic strengths of 1.2 GHz.