The 3D shape of a biological molecule can tell you a lot about what that molecule does – and how its biological activity might be altered, for example to treat a disease.
Scientists at EMBL Grenoble determine 3D structures of human and viral proteins to understand how they interact with the nucleic acids DNA and RNA. To do so, they work closely with instrumentation developers and colleagues across the European Photon and Neutron (EPN) science campus to obtain the best possible data from synchrotron X-ray diffraction or cryo-electron microscopy experiments.
EMBL Grenoble is a key player in the Partnership for Structural Biology (PSB) through which the Grenoble European Photon and Neutron (EPN) Campus institutes – EMBL, the European Synchrotron Radiation Facility (ESRF), the Institut Laue-Langevin, and the Institut de Biologie Structurale (IBS) – provide a uniquely comprehensive range of structural biology platforms for sample production, sample characterisation and structure determination for both in house research and external users.
A cornerstone of the Outstation’s activities is the close interaction with the world-leading ESRF, which produces ultra-intense X–ray beams. EMBL staff collaborate with the ESRF in building and operating state-of-the-art X-ray beamlines, developing associated instrumentation and techniques, and providing expert help to visitors. Thanks to the ESRF Phase I upgrade programme, a new suite of highly sophisticated beamlines is now available for protein crystallography, with fast and automated data collection, and for small-angle X-ray scattering, including a high-throughput sample changer and online high performance liquid chromatography.
High throughput methods have also been introduced in upstream steps of the structure determination process. These include a highly successful robotic platform for nano-volume crystallisation run by the Outstation as well as a recently developed system called CrystalDirect, which enables automated crystal mounting and cryo-cooling. The combination of these technologies with the ESRF massively automated X-ray beamlines has enabled the development of a new generation of fully automated, remote controlled pipelines from protein to X-ray data, streamlining the structural analysis of complex biological targets. They also facilitate structure-guided drug design, through automated facilities for ligand and fragment screening. These platforms are now available to external users under the EU funded iNEXT project. Furthermore, the Outstation has a state-of-the-art Eukaryotic Expression Facility, which features expression of multiprotein complexes in insect cells using the MultiBac technology.
Concerning scientific research, the Outstation focusses on the structural biology of eukaryotic complexes, with a strong tradition in the study of systems involving protein-nucleic acid complexes and viruses. Structural work on aminoacyl-tRNA synthetases is particularly well known and has recently focussed on elucidation of the mode of action of novel boron-containing antibiotics, which target leucyl-tRNA synthetase. Other projects involving protein-RNA interactions include cryo-EM studies of ribosome with the signal recognition particle and translocon and other proteins and complexes involved in RNA processing, transport and degradation, such as the nonsense-mediated decay (NMD) pathway. Further important areas include the analysis of mechanisms of transcriptional regulation, including at the epigenetic level. Groups are working on the structural analysis of eukaryotic transcription factor, chromatin-modification and nucleosome assembly complexes as well as elucidating the mechanisms by which piRNAs (small non-coding RNAs) protect the genome and trying to uncover the role of long non-coding RNAs (lncRNAs). Another focus is the study of segmented RNA viruses, particularly influenza and bunyaviruses, with the aim of understanding how their polymerases replicate and transcribe the viral genome. Complementary to this are studies on the innate immune receptors, which detect the presence of viral RNA in infected cells and activate interferon production.
Scientists at EMBL Grenoble have, via the PSB, access to a wide range of techniques, including molecular biology and biophysical instrumentation, negative stain and cryo-electron microscopy – including a top-end Polara electron microscope with K2 direct detector, and an F20 and T12 –, isotope labelling, nuclear magnetic resonance, neutron scattering, X-ray crystallography and small angle scattering. A confocal microscope with facilities for cross-correlation spectroscopy is available for live cell imaging.
With specialist research groups and teams in both scientific areas research at EMBL Grenoble outstation focuses on structural biology and molecular cell biology. In addition, a number of technology-focused instrumentation teams provide an invaluable resource of technical know how and support to aid the scientific community in the structural biology realm.
Scientists in this unit use integrated structural and computational techniques to study biology at scales from molecular structures to organismal communities.
At its sites in Hamburg
and Grenoble, EMBL provides its researchers and hundreds of external users each year with access to world-leading sources of X-ray and neutron radiation, enabling them to study the structures of biological molecules.