Techniques

Our lab uses cryo EM in combination with single particle reconstruction and fitting of atomic structures to investigate the structural basis of the AAA+ motor activity. For low-resolution single particle EM we use a Technai 120kV microscope equipped with an Eagle 2K CCD camera. High throughput and high resolution microscopy is done using a 300 kV Titan Krios transmission microscope fitted with an Eagle 4K CCD camera. Find a description of the Gene Centres’ state-of-the-art CRYO-ELECTRON MICROSCOPY FACILITY on the webpage of the Beckmann group. We use SPIDER, IMAGIC, EMAN and MRC software packages for image processing on our 4-processor workstations and/or the Linux cluster.

We aim to integrate a variety of structural, biochemical and biophysical methods in order to analyse the dynamic behaviour of motor complexes. Our biochemical expertise comprises standard protein expression and purification methods from E. coli as well as isolation and characterisation of multiprotein AAA+ assemblies from yeast. Where necessary, we will employ Surface Plasmon Resonance, FRET, microscale thermophoresis or SAXS experiments.