Transcript No Slide Title

Supported by:
Daresbury Laboratory
North West Structural Genomics Centre
http://www.nwsgc.ac.uk/
Introduction
Mycobacterium Tuberculosis (TB)
Structural Genomics is a new and rapidly growing interdisciplinary research aimed at extending the vast array of genomic
Tuberculosis kills 2 million people each year. The global epidemic is growing and becoming more dangerous. The
sequence data with a comparable, systematic database of protein structures. Synchrotron Radiation based X-ray Crystallography
breakdown in health services, the spread of HIV/AIDS and the emergence of multidrug-resistant TB are contributing to
is unique in providing very accurate high resolution structures of proteins and their complexes. Thus, translating genome
the worsening impact of this disease.
sequence to large numbers of protein structures via high throughput approaches is of urgent and strategic importance for
* 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
Data
projects they are currently undertaking and have a long term interest in.
Collection
healthcare as well as for fundamental biology.
Cryo
In early 2000, a consortium of several groups in the North West of England
SRS
Preservation
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
On-line Data
Analysis
Crystallisation
and thus initiated the structural genomics effort in the UK. In
Researcher
Target
Protein
Status
Samar Hasnain
Rv0185
Rv2547
Rv2865
Rv0359
Rv2776c
Rv0247c
Rv2718c
Hypothetical metalloprotein
Hypothetical metalloprotein
Hypothetical metalloprotein
Hypothetical metalloprotein
Probable oxidoreductase
Probable Iron-sulphur protein
Probable metalloprotein
Crystallising
Crystals
Cloned (II)
Ligation
Purified
Cloned (I)
Purified
John Helliwell
Rv0510
Rv3307
hemC, porphobilinogen deaminase
deoD, purine nucleoside phosphorylase
Targeted
Targeted
Jordi Bella
Rv0171
Rv1693
Rv1942c
Rv2305
Rv3070
Part of mce1 operon
Hypothetical protein
Conserved hypothetical protein
Hypothetical protein
Unknown membrane protein
Ligation
Ligation
Ligation
Ligation
Ligation
Colin Reynolds
Rv3852
Rv2986c
Rv1388
Rv1407
Histone like protein
Histone like protein
Integration host factor
Similar to other Fmu proteins
Targeted
Cloned (I)
Cloned (I)
Targeted
Lydia Tabernero
Rv0153c
Rv0505c
Rv1967
Rv2234
Rv3042c
Rv3628
Rv3867
Putative tyrosine-phosphatase
serB, probable phosphoserine phosphatase
part of mce3 operon
ptpA, tyrosine-phosphatase
serB2, phospherine phosphatase
ppa,inorganic phosphatase
conserved hypothetical protein
Ligation
Cloned (I)
Ligation
Cloned (I)
Ligation
Cloned (I)
Expressed
Mark Ellis
Rv2060
Rv2229c
Rv2711
Rv3207c
Rv3836
Conserved hypothetical protein
Putative zinc metalloprotein
ideR, iron dependent repressor
Putative zinc metallopeptidase
Putative zinc metallopeptidase
Ligation
Cloned (I)
Purified
Purified
Crystallising
Michele Cianci
Rv3717
Rv3915
Rv2981c
Rv3712
Involved in cell biosynthesis
Involved in cell biosynthesis
Involved in cell biosynthesis
Involved in cell biosynthesis
Targeted
Targeted
Targeted
Targeted
Miroslav Papiz
Rv3910
Rv0588
Rv2154c
Rv2938
Rv1273c
Rv2154c
Membrane protein
Part of mce2 operon
ftsw membrane protein
Transmembrane protein
ABC transporter
Membrane protein
Targeted
Cloned (I)
Cloned (II)
Expressed
Cloned (I)
Cloned (I)
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)
Purification
Phasing
funded a 5 year grant entitled "NW STRUCTURE GENOMICS
CENTRE'S HIGH THROUGHPUT MAD BEAMLINE FOR
PATHOGENS GENOMES”. The grant entitles the NWSGC 67%
of the beam time.
Expression
Modelling
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
BL 10 Exp. Hutch
Insertion Device
Monochromator
DNA
Structure
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'’.
Targets
High Throughput
Protein Structure
Determination
Drug
Design
Comparison of MAD 10 and ESRF BM14
MAD Beamline 10 @ The SRS
MAD MPW Output
1.4E+14
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.
1.2E+14
Flux (photons/s/mrad/250mA/0.1%)
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.
2.4 T, 9 Pole MPW
ESRF BM 14
1E+14
BL10 2.4 T
8E+13
6E+13
4E+13
ESRF BM14
2E+13
0
4
8
12
Photon Energy (keV)
16
20