Fast High-Resolution Protein Structure Determination by Using Unassigned NMR Data

摘要

NMR spectroscopy provides high-resolution structural information of biomolecules in near-physiological conditions. Although significant improvements were achieved in NMR spectroscopy in the last 20 years,[1] the increase in genome sequencing data has created a need for rapid and efficient methods of NMR-based structure determination.[2], [3] NMR data acquisition can be accelerated significantly when sensitive spectrometers are combined with new methods for sampling chemical shifts in multidimensional NMR experiments.[4] Therefore, data analysis and in particular the requirement to assign side-chain chemical shifts to specific atoms is the major bottleneck of rapid NMR-based structure determination. Herein, we present a method, termed FastNMR (fast structure determination by NMR) that enables automatic, high-resolution NMR structure determination of one-domain-sized proteins from unassigned NMR data. By using FastNMR, the de novo structure of the 65-residue cone snail neurotoxin conkunitzin-S2 was determined automatically.