Fast protein backbone NMR resonance assignment using the BATCH strategy.

Bernhard Brutscher , Ewen Lescop (1)

1 ICSN - Institut de Chimie des Substances Naturelles

Probing protein structure, dynamics, and interaction surfaces by NMR requires initial backbone resonance assignment. The protocol for this step has been progressively developed in the last 15 years to provide robust assignments. However, even in the case of favorable conditions (high field magnets and cryogenically cooled probes, small globular proteins, high sample concentration), the assignment step generally takes several days of data collection and analysis, thus precluding studies of unstable proteins and limiting high-throughput applications. Recently, we have introduced the BATCH strategy for fast protein backbone resonance assignment. BATCH benefits from the combination of several tools (BEST/ASCOM/Targeted-Sampling/COBRA/HADAMAC) for time-optimized and highly automated NMR data acquisition, processing, and analysis. In this chapter, we discuss the individual steps of the BATCH method and describe its practical implementation to obtain the backbone resonance assignment of small globular proteins in a few hours of time.

Domaines: Chimie organique

Bernhard Brutscher, Ewen Lescop. Fast protein backbone NMR resonance assignment using the BATCH strategy.. Methods in Molecular Biology, 2012, 831, pp.407-28. ⟨10.1007/978-1-61779-480-3_21⟩. ⟨hal-00798115⟩