- Institut Pasteur, CNRS URA 2185  


  HEADDr. HAOUZ Ahmed / ahaouz@pasteur.fr
  MEMBERSFRAYSSE Jocelyne / MIRAS Isabelle / NAVAZA Rafael/ Dr. SHEPARD William / WEBER Patrick


  Annual Report

The goal of the platform 6 (PF6) is to provide 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 during the period (2001-2007). New or upgraded equipment includes a complete X-ray diffraction facility (MicroMax007 rotating anode generator, MarResearch 345dtb Image Plate detector, Oxford cryogenic system), three automatic workstations for protein crystallization (robot TECAN Genesis150, Cartesian Technologies nanodispenser and Matrix Maker ), a microscope with motorized plate (Nikon Eclipse E600) for crystal visualization, and central computing facilities for crystallography and LIMS databases.

In the case of our service facility, during 2008, more than 500 different proteins sample 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 samples (about 80%) originated from different crystallographic teams of the Institut Pasteur.

The PF6 also participates in several other research projects (PTR, ANR,....) involving structural studies of single proteins or protein complexes. These projects arise from direct collaborations of the PF6 with biology labs at the Institut Pasteur and outside research organisations. During 2008, we have obtained more than ten diffracting crystals and solved five structures by using SAD or MAD techniques on SeMet-labeled proteins.

Keywords: Protein crystallography, Structural genomics, automated crystallogenesis, X-ray diffraction, tuberculosis, leprosy

Ptcristallo.jpg

Figure: Crystallization and X-ray diffraction facilities present in Platform 6



  Publications

Bellinzoni M, Haouz A, Grana M, Munier-Lehmann H, Shepard W, Alzari PM.(2006). The crystal structure of Mycobacterium tuberculosis adenylate kinase in complex with two molecules of ADP and Mg2+ supports an associative mechanism for phosphoryl transfer. Protein Science15:1489-93, PMID: 16672241.

Geerlof A, Brown J, Coutard B, Egloff MP, Enguita FJ, Fogg MJ, Gilbert RJ, Groves MR, Haouz A, Nettleship JE, Nordlund P, Owens RJ, Ruff M, Sainsbury S, Svergun DI, Wilmanns M.(2006). The impact of protein characterization in structural proteomics. Acta Crystallogr D Biol Crystallogr. 62 :1125-36. PMID: 17001090.

Delarue M; Duclert-Savatier N, Miclet E, Haouz A,Giganti D; Ouazzani J, Lopez P, Nilges, and M; Stoven V . (2007). Three dimensional structure and implications for the catalytic mechanism of 6- phosphogluconolactonase from Trypanosoma brucei. Journal of Molecular Biology 366 : 868-881. PMID: 17196981

Shepard W, Haouz A, Grana M, Buschiazzo A, Betton J-M, Cole ST, Alzari PM. The crystal structure of Rv0813c from Mycobacterium tuberculosis reveals a new family of FABP-like proteins in bacteria. Journal of Bacteriology, 189: 1899-1904. PMID: 17172346

Barbey C, Rouhier N, Haouz A,Navaza A, Jacquot JP. Overproduction, purification, crystallization and preliminary X-ray analysis of the peroxiredoxin domain of a larger natural hybrid protein from Thermotoga maritima. (2008). Acta Crystallogr Sect F Struct Biol Cryst Commun. 64 : 29-31. PMID: 18097097



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Activity Reports 2009 - Institut Pasteur
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