Top-down strategies, based on the study of entire proteins, hold great promise for structural protein analysis. Whilst the classical, bottom-up approach can be useful for locating proteins already deposited in the databases, only a top-down methodology can yield complete and unambiguous structural characterization in a time efficient manner.
In this regard a particularly important application is the analysis and localisation of post-translational modifications. This often relies on efficient, selective and comprehensive fragmentation such as that provided by electron capture dissociation (ECD) or Electron Transfer Dissociation (ETD).
Most recently these techniques have been used in the analysis of PilE, an 18 kDa protein that is strongly involved in the virulence of Neisseria Meningitidis.
Our unit is part of the recent consortium for Top-Down Proteomics.
To improve the crucial stage of detection and characterisation of cross-linked peptides, we have developed a new generation of cross-linking agents based on click-chemistry.
This work is performed in collaboration with the group of Prof. Pierre-Yves Renard from the University of Rouen, who is responsible for the synthesis of the novel cross-linkers and also the group of G. Duménil (Hôpital Eurpéen Georges Pompidou) for all biological aspects involving type IV pili of N.meningitidis.
This project is funded for 2009-2013 thanks to an ANR PIRIBio grant.