- (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.
Mancini EJ, Kainov DE, Grimes JM, Tuma R, Bamford DH, Stuart
DI. Cell. 2004 Sep 17;118(6):743-55.
- (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that allowed us to sail through the commissioning time, has been replaced by a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
|
l
(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
|
First
test images were obtained on
14
November 2003
on a motor neuron causing mutant of human superoxide dismutase with a cell
dimension (a=166.16,
b=203.68, c=144.48Å). Data went to 1.8Å Resolution and reasonable images were
obtained in 45 Seconds for 0.5° oscillation.
-
(31st of October 2003) First crystallographic tests
Tests made by Manchester University Structural
Chemistry Laboratory staff (J R and M Helliwell with M Cianci) on
31st October 2003
with a small molecule crystal showed that the precision of the instrument
wavelength setting and geometric reproducibility of the line 10 camera allowed
correct diffraction pattern prediction.
-
(8th/9th of September 2003) SRS User Meeting Poster: Status
of NWSGC MAD10
-
(11th of August 2003) First XAFS data collected off BL10 Monochromator
-
|