Methods

Passmore Lab

 

Hybrid approach

We often start by purifying native protein complexes from the yeast Saccharomyces cerevisiae, using TAP tagging approaches, to determine their structures and characterise their functions. Most projects have then moved onto recombinant protein expression in insect cells. The techniques we use include electron cryo-microscopy (cryo-EM), as well as x-ray crystallographic, biophysical, biochemical and genetic methods. 


Recent developments in electron microscopes, direct electron detectors and software have brought about a revolution in cryo-EM. Near-atomic resolution structures are now possible. My lab is developing new methods to further improve images. The MRC Laboratory of Molecular Biology electron microscopy facility houses a number of FEI microscopes, including two 300 keV Titan Krios each with two direct electron detectors and energy filter, a 300 keV Polara (also with a direct electron detector and energy filter) and microscopes for sample preparation.


 

Passmore Lab, MRC Laboratory of Molecular Biology, Cambridge UK