Márquez Team
The high throughput crystallisation lab at the PSB
High-throughput crystallisation robot at the HTX Lab.
Structure of the Abscisic acid hormone receptor showing the gating loops in the closed (magenta) and open (green) conformations.
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 newmethods and concepts in macromolecular crystallography and works in close coordination with the high-throughput protein expression and synchrotron instrumentationgroups 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) formacromolecular crystallography.We are also working in collaboration with the outstation’s Instrumentation Group (page90) 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.