Is NMR slow? From gene to fold in only two weeks

RASIA RM

Tucumán

Congreso; XLV Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2009

The identification of protein constructs amenable to structural characterization is currently a bottleneck in biomolecular NMR. Studying biological assemblies often involves optimizing each component separately and reforming the complex in vitro. Likewise, investigating the structure and function of modular proteins can require each domain being excised and studied independently.We have set up a simple method for parallel expression, labeling and purification of protein constructs (up to 80 kDa) combined with rapid evaluation by NMR spectroscopy. Our approach, which is equally applicable for manual or automatic implementation, offers an efficient way to identify and optimize protein constructs for NMR. RDC restraints are then obtained for each construct using optimized liquid crystal NMR experiments and analysis tools. These data sets are used directly to calculate the protein fold using the Rosetta NMR software.