Workpackage 7

Streamlining Access Procedures for Pioneering Structural Biology Methods

Here we are focusing on streamlining procedures for several rare or unique technologies available through Instruct that have been opened up for general access to the structural biology community. The increased demand requires that we develop the level of standardisation and robustness consistent with expected levels of user access. In Task 7.1, procedures for methyl specific labelling will be established that allow in vitro and in cell analyses of large molecular complexes by NMR, permitting direct correlation with results obtained on the same sample by electron microscopy. In Task 7.2, we will develop a work flow to improve the efficiency of production of integral membrane proteins, normally a slow trial-by-error manual process. Targets will enter this work flow and proceed through a structured protocol where various parameters are tested in logical stepwise manner in bacterial, insect and mammalian cells. In Task 7.3, the repertoire of applications for nanobodies will be increased, thereby widening its user base. Nanobodies have already shown huge potential as crystallisation chaperones for membrane proteins. Here we will simplify the isolation of binders from nanobody phage display libraries and establish new applications as intracellular fluorescent biosensors.