News 2004
October 2004
Meeting Science Minister Lord Sainsbury
Staff scientist Michele Cianci, of NWSGC Structural Biology beamline MAD 10,
met Lord Sainsbury recently and informed him of the latest performance of this
world competitive beamline at the SRS at Daresbury.
October 2004
NWSGC MAD10 beamline contributes to structure solution of ESCRT-II,
an Endosome-Associated Complex Required for Protein Sorting
Dr. Roger L. Williams from LMB-MRC (UK)
recently reported in Developmental Cell the crystal structure ofESCRT-II,
an Endosome-Associated Complex required for Protein Sorting. Essential contributions
for model building come from data collected at NWSGC MAD10 beamline at Daresbury
during commissioning time.
For more information please click here.
September 2004
Beamline 10 in the SRD Annual Report 2003-2004
The progress of the development of beamline 10 is well
documented in the SRD
Annual Report 2003-2004, page 10 (of course...)
September 2004
First Structure solved By MAD
Dr. Fusinita van den Ent and Dr. Jan Lowe (LMB-MRC)
were the first users to solve a structure using the MAD capability of SRS beamline
10.1. They collected data overnight on Friday and by the evening of the following
Sunday they were able to state 'The structure is basically solved. The native
is 2.8 Å, we couldn't get anything better, but that should be enough.
The density is gorgeous!'
September 2004
First paper published for work carried out on beamline 10
The protein of interest (structure already solved) binds
NTPs and Mg. Mancini et al. aimed to collect protein+ATP+Mg and protein+ATP+Mn
to verify the Mg position by collecting data near the Mn K edge. Datasets were
collected using X-rays of 1.8A and showed clear (20 sigma) peaks in the anomalous
maps. The second dataset showed weak Mg anomalous sites at the same places.
August 2004
Installation of MAR225 CCD Detector
In August 2004 the glorious MAR165 CCD from MAR-USA that
allowed us to sail through the commissioning time, has been replaced by a 3x3
tiled MAR 225 CCD detector. This has an active area of 225x225mm2 with
pixel size of 73 microns. Readout time is 1 second with 3.5 seconds of dead
time. In the photograph Dr. Lentfer, Dr. Blum and Prof. Hasnain during
the installation.
April 2004
Installation and commissioning of Fluorescence detector
The optimum wavelength for MAD experiment is provided by the fluorescence data
obtained from a monolithic low profile Ge detector, which also provides capability
of collecting XAFS data on the same crystals as well as monitor the redox state
of a ‘metallic’ functional group during crystallographic data collection.
Figure below shows a close in view of the sample area with the monolithic fluorescence
detector and automatic sample changer.
March 2004
First MAD experiments
Dr. Andrew Sharff from Astex Technology recently collected a couple
of MAD data sets (at the Hg LIII edge) on a current drug target as part of benchmarking
efforts of the new beamline. These have provided the heavy atom site and Andrew
is hopeful “that structure solution would soon emerge." The data went
to 2.5Å resolution (cell 60 x 50 x 103 Å – space group P21).
March 2004
More testings ...
A
group from AstraZeneca had similar experience. Dr. Julie Tucker and Stefan
Gerhardt from AstraZeneca collected a couple of data sets on a ligand bound
complex of a phosphorylase on NWSGC MAD 10 as part of benchmarking effort of
the new beamline. Julie and Stefan were able to extend the resolution (unit
cell size 115, 127, 127Å, P212121) to 1.9Å compared to data collected
on 14.2, where very similar crystals diffracted to 2.3Å.
February 2004
Pushing resolution limits ...
Further tests on thrombin crystals (provided in a collaboration with
GSK) in February 2004 assessed the diffraction resolution limits and showed
comparable results to an undulator insertion device based instrument whereby
good data up to 1.4Å were measured on line 10.
February 2004
First data collection on crystals with large unit cell and high resolution
Table below provides the summary of two data sets obtained for two forms of
crystals of the same protein, human superoxide dismutase. These data were collected
by Drs. Michele Cianci and Svetlana Antonyuk. One of these is for a reasonable
modest cell dimensions (the largest axis being 203Å) and the other representing
a relatively smaller cell (the largest axis being 67Å). The atomic resolution
achieved in the latter case and high resolution achieved in the former case
provided confirmation that beam line is able to provide high quality data.
| C2221 | P21 | |
| 1(Å) | 1.37 | 1.07 |
| Name of the sample | HSOD | HSOD |
| Resolution range (Å) | 50-1.95 (2.02-1.95) |
50-1.24 (1.28 - 1.24) |
| Completeness (%) | 86.4 (90.8) | 96.3 (82.5) |
| R merge | 6.5 | 6.3( 41.0) |
| <I>/sI last shell | 2.35 | 2.0 |
| Redundancy | 8 | 4 |
| Overall reflections | 1341091 | 1116141 |
| Unique reflections | 152998 | 70055 |
| Wilson B-factor (Å2) | 30 | 12 |
Unit cell parameters a b c |
166.18 203.59 145.58 90° 90° 90° |
38.6 67.5 52.3 90° 106.5° 90° |
| Solvent (%) | 68 | 40 |
| Number of dimers | 5 | 1 |
