Crystallogenesis and X-Ray Diffraction


  HEADDr HAOUZ Ahmed / ahaouz@pasteur.fr
  MEMBERSBENHARRATS Hind / NAVAZA Raphael


  Annual Report

The goal of the platform 6 is to provide the research teams working on macromolecular crystallography at Pasteur with the equipment and technology required for automated crystallogenesis, computing and X ray diffraction measurements, and develop high-throughput methodologies for gene cloning, protein production and crystallization by automating the different steps of the process for structural genomics studies. Funding from the Genopole®Ile de France, the French Ministry of Research, the European Commission and the Pasteur Institute has allowed us to make significant investments in equipment for the project on the structural genomics of mycobacteria (2001-2006). New or upgraded equipment includes a complete X-ray diffraction facility (MicroMax007 rotating anode generator, MarResearch 345dtb Image Plate detector, cryogenic systems), two automatic workstations for protein crystallization (robot TECAN Genesis150, Cartesian Technologies nanodispenser), a microscope with motorized plate (Nikon Eclipse E600) for crystal visualization, and central computing facilities for crystallography and LIMS databases. Our latest acquisition is a Matrix Maker automatic workstation (Emerald BioSystems) for crystal optimization that should be operational in early 2007.

At present, more than 200 different proteins have been submitted to automatic crystallization assays, with a success rate (diffracting crystals or exploitable hints) of roughly one of every three proteins assayed. Most of these proteins (about 60%) originated from the Structural Genomics of Mycobacteria project piloted by Prof. Pedro ALZARI (IP), As of September 2006, 535 mycobacterial genes have been cloned into bacterial expression vectors using Gateway or related technologies, 175 gene products were expressed as soluble proteins and 75 of these purified to homogeneity and subjected to crystallization trials. We have determined so far 17 crystal structures, mostly using SAD or MAD techniques on SeMet-labeled proteins, and diffraction data has been collected for 5 other proteins for which structure determination is in progress.

The PF6 also participates in several other research projects (PTR, GPH) involving structural studies of single proteins or protein complexes. These projects arise from the crystallography labs or from direct collaborations of the PF6 with biology labs at the Pasteur Institute and outside.

haouz.jpg

Quelques cristaux de protéines à fonction inconnue obtenus dans le cadre du projet de génomique structurale des mycobactéries.

Some crystals of mycobacterial proteins obtained at the PF6.



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More informations on our web site


  Publications

Publications 2006 of the unit on Pasteur's references database




Activity Reports 2006 - Institut Pasteur
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