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 architectures and characterise their functions. We translate these findings to establish recombinant expression of protein complexes 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. It is now possible to build atomic models directly from cryo-EM maps. Recently, the Passmore lab developed new gold and graphene specimen supports that further improve cryo-EM images. The MRC Laboratory of Molecular Biology electron microscopy facility houses a number of FEI microscopes, including three 300 keV Titan Krios each with two direct electron detectors and energy filter, two 300 keV Polaras (also with a direct electron detectors), an F20, and microscopes for sample preparation.



























We ran cryo-EM courses in 2014 and 2017 as an introduction to the method. The lectures are available online from the following link:

         https://www2.mrc-lmb.cam.ac.uk/research/scientific-training/


 

Passmore Lab, MRC Laboratory of Molecular Biology, Cambridge UK