UNDERSTANDING THE MOLECULAR BASIS OF METABOLIC PROTEIN CLUSTERS

Enzymes frequently cluster into large higher-order structures, also termed “metabolons”, to execute sequential, multistep cascade reactions. These macromolecular complexes provide several metabolic advantages, such as substrates channelling between catalytic sites, higher flux rates that are important if the substrate intermediates are instable (i.e. short half-life) and they ensure a high overall catalytic efficiency.
We are using state-of-the-art microscopy methods, such as single particle analysis (SPA), correlative light and electron microscopy (CLEM) combined with FIB-SEM and time-resolved electron microscopy (TREM) to characterise the structure & architecture of such large complex assemblies. We will visualise conformational changes upon substrate binding and determine the underlying kinetics using classical biochemical and biophysical methods. The figure on the right shows the overall architecture of one of our target complexes (negative stain microscopy; top) and the spraying device that will be used to conduct TREM studies (bottom).