ayout 1 - EMBL Grenoble

EMBL Research at a Glance 2009
José A. Márquez
PhD 1997, University of
Valencia.
Postdoctoral research at
EMBL.
Staff Scientist at EMBL
Grenoble since 2003.
Team leader since 2005.
The high-throughput crystallisation laboratory at
EMBL Grenoble
Previous and current research
Finding conditions in which biological macromolecules form crystals is recognised as one of the
major bottlenecks in structural biology. Once macromolecules are purified, they need to be assayed
for crystallisation with a collection of precipitants under different chemical environments. This
leads to the need to perform hundreds of experiments, consuming large amounts of sample and
taking time. At EMBL Grenoble we have established a high-throughput crystallisation platform,
the HTX Lab, with the aim to increase the success rate and speed up the process of crystal structure
determination. In this platform the whole process of crystallisation screening is automated
through the introduction of liquid handling, crystallisation and crystal imaging robots. Starting
in April 2009 the HTX Lab will offer automated crystallisation screening services to European research
through the EC-funded PCUBE project.
The technology introduced allows us to perform experiments using extremely low volumes of
sample, which makes it possible to perform extensive screening even when the amount of sample
is limited. This platform, which started to operate in September 2003, has now more than three
hundred registered users, and over two million individual crystallisation experiments have already been performed. The high-throughput crystallisation
laboratory is not only open to EMBL researchers but also to all the members of the Partnership for Structural Biology (PSB), which
includes the ESRF, the ILL, the IBS and the IVMS, and represents one of its core technological platforms. Access is also granted to European
researchers through the PCUBE project.
Future projects and goals
In addition to offering automated crystallisation resources, the HTX lab is actively involved in the development of new methods and concepts
in macromolecular crystallography and works in close coordination with the high-throughput protein expression and synchrotron instrumentation
groups at the outstation. One of our major areas of development is data management. We are collaborating with the EBI, EMBL
Hamburg, EMBL Heidelberg and other laboratories in Europe towards the development of a common Laboratory Information Management
System (LIMS) for macromolecular crystallography. We are also working in collaboration with the outstation’s Instrumentation Group (page
90) in order to develop strategies to close the gap between crystallisation
and data collection by facilitating operations like
crystal mounting or freezing that are required before data collection.
We are currently applying high-throughput methods to the
study of signalling molecules and transcriptional regulators.
We have recently solved the structure of the extracellular domain
of the human inhibitory receptor IREM-1 expressed in
myeloid cells and we are now investigating other members of
this receptor family.
High-throughput crystallisation
robot at the HTX Lab.
Selected references
Dimasi, N., Flot, D., Dupeux, F. & Marquez, J.A. (2007). Expression,
crystallization and X-ray data collection from microcrystals of the
extracellular domain of the human inhibitory receptor expressed on
myeloid cells IREM-1. Acta Crystallogr. Sect. F Struct. Biol. Cryst.
Commun., 63, 20-208
Marquez, J.A., Galfre, E., Dupeux, F., Flot, D., Moran, O. & Dimasi,
N. (2007). The crystal structure of the extracellular domain of the
inhibitor receptor expressed on myeloid cells IREM-1. J. Mol. Biol.,
367, 310-318
Marquez, J., Reinelt, S., Koch, B., Engelmann, R., Hengstenberg, W.
& Scheffzek, K. (2006). Structure of the full-length enzyme I of the
phosphoenolpyruvate-dependent sugar phosphotransferase system.
J. Biol. Chem., 281, 32508-32515
Marquez, J.A., Smith, C.I., Petoukhov, M.V., Lo Surdo, P., Mattsson,
P.T., Knekt, M., Westlund, A., Scheffzek, K., Saraste, M. & Svergun,
D.I. (2003). Conformation of full-length Bruton tyrosine kinase (Btk)
from synchrotron X-ray solution scattering. EMBO J., 22, 616-62
92