ARIA will be unavailable on 24th February 2025 due to scheduled maintenance. Please plan work accordingly.
Instruct PartnerRegistration is now open for the next FRISBI/ReNaFoBis webinar
Speaker: Nils Marechal, IGBMC/CBI, Illkirch
Tittle : Streptavidin affinity grids for structural electron microscopy
Summary: Despite considerable technological and methodological breakthroughs, the main limitation impairing structure determination by cryo-EM remains the electron microscopy specimen preparation. During the standard plunge-freezing procedure, the biological sample undergoes mechanical stress including shearing forces and experiences the close proximity of liquid-gas interfaces. In many cases, these physico-chemical conditions can lead to sample denaturation, dissociation, aggregation or anisotropic angular distribution. A variety of approaches were proposed to overcome these limits such as the development of high-speed plunge-freezing system, supplementation of the sample buffer with detergents to block interfaces or the use of thin support films. I will present different strategies based on affinity layers which not only allow to sequester the sample far from disruptive interfaces, but also permit to work with a reduced sample amount and to tightly control the sample behavior. We will focus on one of these approaches: the streptavidin grids. Streptavidin affinity grids promise to be of particular interest to study machineries involved in gene expression regulation thanks to the possibility to readily capture the macromolecular targets through specific interactions with a biotinylated nucleic acid. With Lamour team at IGBMC, we recently used this approach to solve the first structure of E. coli DNA Gyrase bound to a DNA crossover on a DNA minicircle.
PLEASE REGISTER HERE: https://us06web.zoom.us/webinar/register/WN_r7YokBr5Sp-CH6AEm0v7hQ
