Mycobacterium Tuberculosis (TB)

1. Daresbury Laboratory BL10 2.4 T ESRF BM14 Phasing Monochromator BL 10 Exp. Hutch Insertion Device Supported by: North West Structural Genomics Centre http://www.nwsgc.ac.uk/ Introduction Structural Genomics is a new and rapidly growing interdisciplinary research aimed at extending the vast array of genomic sequence data with a comparable, systematic database of protein structures. Synchrotron Radiation based X-ray Crystallography is unique in providing very accurate high resolution structures of proteins and their complexes. Thus, translating genome sequence to large numbers of protein structures via high throughput approaches is of urgent and strategic importance for healthcare as well as for fundamental biology. In early 2000, a consortium of several groups in the North West of England proposed the establishment of a structure genomics centre (NWSGC). The NWSGC members have brought together expertise in X-ray protein crystallography, pathogens biology, membrane proteins, metalloproteins and thus initiated the structural genomics effort in the UK. In summer 2001 Leeds University Bioinformatics group joined the NWSGC, followed by Astex Technology of Cambridge in 2002. In July 2001, UK's research councils (BBSRC, EPSRC & MRC) funded a 5 year grantentitled "NW STRUCTURE GENOMICS CENTRE'S HIGH THROUGHPUT MAD BEAMLINE FOR PATHOGENS GENOMES”. The grant entitles the NWSGC 67% of the beam time. Our long-term aim is to form links with other genomics efforts in the UK. NWSGC has selected to join the International TB effort and has established close links with the RIKEN structural genomics programme. In June 2002, we have participated in two EOI’s for EU framework VI proposal; one entitled ‘Tuberculosis Drug Development’ and the other entitled ‘Structural Genomics of Metalloprotein 'Function and Mis-function'’. Mycobacterium Tuberculosis (TB) Tuberculosis kills 2 million people each year. The global epidemic is growing and becoming more dangerous. The breakdown in health services, the spread of HIV/AIDS and the emergence of multidrug-resistant TB are contributing to the worsening impact of this disease. * Nearly one percent of the world's population is newly infected with TB each year. * Overall, one-third of the world's population is currently infected with the TB bacillus. * 5 - 10 percent of people who are infected with TB become sick or infectious at some time during their life. The members of the NWSGC have chosen targets from the TB genome which relate to projects they are currently undertaking and have a long term interest in. Cryo Preservation SRS Data Collection Researcher Target Protein Status Samar Hasnain Rv0185 Hypothetical metalloprotein Crystallising Rv2547 Hypothetical metalloprotein Crystals Rv2865 Hypothetical metalloprotein Cloned (II) Rv0359 Hypothetical metalloprotein Ligation Rv2776c Probable oxidoreductase Purified Rv0247c Probable Iron-sulphur protein Cloned (I) Rv2718c Probable metalloprotein Purified John Helliwell Rv0510 hemC, porphobilinogen deaminase Targeted Rv3307 deoD, purine nucleoside phosphorylase Targeted Jordi Bella Rv0171 Part of mce1 operon Ligation Rv1693 Hypothetical protein Ligation Rv1942c Conserved hypothetical protein Ligation Rv2305 Hypothetical protein Ligation Rv3070 Unknown membrane protein Ligation Colin Reynolds Rv3852 Histone like protein Targeted Rv2986c Histone like protein Cloned (I) Rv1388 Integration host factor Cloned (I) Rv1407 Similar to other Fmu proteins Targeted Lydia Tabernero Rv0153c Putative tyrosine-phosphatase Ligation Rv0505c serB, probable phosphoserine phosphatase Cloned (I) Rv1967 part of mce3 operon Ligation Rv2234 ptpA, tyrosine-phosphatase Cloned (I) Rv3042c serB2, phospherine phosphatase Ligation Rv3628 ppa,inorganic phosphatase Cloned (I) Rv3867 conserved hypothetical protein Expressed Mark Ellis Rv2060 Conserved hypothetical protein Ligation Rv2229c Putative zinc metalloprotein Cloned (I) Rv2711 ideR, iron dependent repressor Purified Rv3207c Putative zinc metallopeptidase Purified Rv3836 Putative zinc metallopeptidase Crystallising Michele Cianci Rv3717 Involved in cell biosynthesis Targeted Rv3915 Involved in cell biosynthesis Targeted Rv2981c Involved in cell biosynthesis Targeted Rv3712 Involved in cell biosynthesis Targeted Miroslav Papiz Rv3910 Membrane protein Targeted Rv0588 Part of mce2 operon Cloned (I) Rv2154c ftsw membrane protein Cloned (II) Rv2938 Transmembrane protein Expressed Rv1273c ABC transporter Cloned (I) Rv2154c Membrane protein Cloned (I) On-line Data Analysis Crystallisation Purification Expression Modelling DNA Structure High Throughput Protein Structure Determination Drug Design Targets Comparison of MAD 10 and ESRF BM14 MAD Beamline 10 @ The SRS The new 2.4 tesla 9 pole wiggler beamline (MPW10) has been developed by Daresbury project team staff from ASTeC, ED and SRD. The optical arrangement is optimised through a 100 micron collimator and for rapid tuneability and high energy resolution allowing data to be collected from small, weakly diffracting crystals over a wide range of wavelengths. It will be dedicated to MAD structure solution operating in the 4 to 14 keV photon energy range. A program of development will be undertaken with the aim of making the protein crystallography facilities as easy to use as possible. This includes automated sample changers, on line data processing and remote operation. The performance of NWSGC MAD 10 will be in the same order of magnitude of the very successful ESRF BM14. These characteristics are required for high throughput protein crystallography, where samples of unknown quality will arrive for screening and immediate data collection.