IBR   13079
INSTITUTO DE BIOLOGIA MOLECULAR Y CELULAR DE ROSARIO
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
A fast strategy for structural characterization of multidomain proteins. Application to HYL1-RNA intaraction. (charla por invitación)
Autor/es:
RASIA RM; BOLOGNA NG; NOIRCLERC-SAVOYE M; GALLET B; VERNET T; BRUTSCHER B; PALATNIK JF; BOISBOUVIER J
Lugar:
Murnau, Alemania
Reunión:
Workshop; Advances in NMR of protein and nucleic acide molecular recognition; 2008
Resumen:
A major challenge in structural biology remains the identification of protein constructs amenable to structural characterization. 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. The backbone of the selected constructs are then assigne and RND restraints are obtained using optimized liquid crystal NMR experiments and analysis tools. These datasets are used directly to calculate the protein fold using Rosetta NMR software.Several multidomain proteins are known to participate in microRNA processing in plants. Here we show an application of this procedure to the structural characterization of the multidomain protein HYL1 from A. thaliana and of its interaction with RNA substrates.