|mosquito® Crystal and mosquito® LCP: fast, reliable automation of protein crystallisation drop set-up |
Joby Jenkins, David Smith
This poster demonstrates the ability of TTP Labtech’s mosquito® Crystal and mosquito® LCP to address the issues inherent in the automated set-up of protein crystallisation screen trials, without needing constant instrument configuration changes.
|Structure determination of the virulence factor AsaP1 from Aeromonas salmonicida subsp. Achromogenes|
Xenia Bogdanovic, Rajesh Kumar Singh, Gottfried J. Palm, Johanna Hentschke, Bjarnheidur K. Gudmundsdóttir & Winfried Hinrichs
Aeromonas commonly found in waters, wastewaters and mud are non-motile gram negative bacteria. Atypical strains of Aeromonas salmonicida are known to be largely responsible for causing a lethal disease in Atlantic salmons and other farmed fishes.
|Stable N-phosphorylated triazol-5-ylidenes|
Kirilchuk,A. A.; Marchenko,A. P.; Koidan, H. G.; Zarudnitskii, E. V.; Huryeva, A. N.; Yurchenko, A. A.; Kostyuk, A. N.
Considering probable generality of “carbene” mechanism for 1,3-azoles, investigation of intermediates of phosphorylation of 1,2,4-triazoles is of interest.
|A Crystallographic-Based Fragment Screen Against Human BRD4 Bromodomain 1|
Hubbard P, Ritchie A, Hughes S, Lipkin M, Blackaby W & MacLeod A
Fragment-based drug discovery has now become firmly established as an important method in the lead discovery process, and is an integral component of drug discovery at BioFocus. Using a selection of compounds from our fragment collection, we have performed a crystallographic-based screen against human BRD4 bromodomain 1.
|A fast and fully automated solution for Lipidic Cubic Phase (LCP) screening using mosquito LCP|
Joby Jenkins1, Patricia Edwards2, Rob Lewis1and Joanne Franklin1
Membrane proteins, such as G-protein-coupled receptors, are known to be much more difficult to purify and crystallise than soluble proteins due to their native environment within the lipid bilayer of the cell membrane. The in meso (lipidic cubic phase or LCP) crystallisation technique has revolutionised the process of crystallising membrane proteins. This method utilises highly viscous lipid mesophases to contain the membrane proteins for crystallisation.
|mosquito® Crystal: Fast, reliable automation of Protein Crystallization drop set-up|
Joby Jenkins, David Smith, Chloe Carter, Wendy Gaisford
Automation of protein crystallography screening has contributed significantly to the rapid progress of crystallography based structural biology. Automation allows samples to be screened using smaller volumes of both protein and screen solutions, reducing costs and saving valuable protein. Additional benefits include increased throughput and accuracy. One of the challenges to automating this process is the necessity to accurately pipette solutions of varying viscosities
|Analysis of overall success of robotic crystallization|
I. Ivanovic, H. M. Baker, J. S. Lott and E. N. Baker
A total of about 140 different proteins have been commissioned and 62 have been crystallized with a quality suitable for X-ray diffraction (45%). On a per-drop basis, 300 crystallization experiments have been successful, representing an over¬all success rate of 3%. PEG 3350 was the precipitant for 20, making it the single most successful crystallization agent.
|Strategies for expression and solubility analysis|
Production of soluble protein is one of the major bottlenecks that precede crystallographic studies.During the last years several techniques and strategies have been developed to address this problem. However, many of them imply an economical cost and technologies that are not always available. We will describe a general plan for protein solubility analysis by using a combination of four different but complementary strategies. In this plan, different constructs of a protein interest are designed
|Automating Microseeding Protein Crystallography Set-ups Using Mosquito|
Joby Jenkins, David Smith, Rob Lewis, Chloe Carter
Crystallising proteins, required for structure determination by X-ray diffraction, is a difficult and labour-intensive task. The mosquito liquid handler (TTP LabTech) is ideally suited to automating the complex set-ups required for microseeding due to its precide handling of extremely low volumes of even viscous solutions, and its ability to perform multiple aspirations and dispenses with each pipette.
|Automated Single Crystal Structure Determination - A Tool for Synthetic Chemists?|
Bernd Hinrichsen, Martin Adam, Michael Carr, Dieter Schollmeyer,
During recent years large improvements in software functionality and its ease-of-use have made single crystal X-ray structure determination easier than ever. These days most structures can be measured, processed, solved and refined using well selected defaults with no or little crystallographic knowledge. Recently, microfocus sources and CCD detectors both air-cooled, have entered the marketplace. Combining these innovations with an automated sample loader and an intelligent graphical user inte
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