Structural Biology

Proteins and complexes of proteins with other macromolecules such as RNA/DNA or carbohydrates are the functional units underlying all cellular activities. To understand cellular function and dysfunction in human health and disease, a description of the structure and function of individual proteins and protein complexes is indispensable for basic, as well as translational, science.

The Structural Biology Science Technology Platform (SB STP) comprises a team of skilled scientists who provide gated access to world-class facilities, expertise and support in the complementary areas of specialised protein expression and purification, biophysical analysis and characterisation, protein crystallisation and structure determination (principally by X-ray crystallography, but supported where necessary by other methods such as SAXS) and lab automation.

The protein production facility within the SB STP draws upon a solid body of experience to support collaborators at various stages of the procedure from protein-coding DNA sequence to recombinant protein.

The facility has a vast collection of annotated vectors and associated sequencing primers to get collaborations with users swiftly off the ground and also provides in-house vector design, troubleshooting and training. Depending on the nature of the protein/project, the facility can offer expression in E.coli, insect (baculovirus) or mammalian cells (transient expression). High-throughput cloning using Ligation Independent Cloning (LIC), protein domain boundary definition (design, clone and express on a small scale up to 96 constructs) and solubility screening (affinity-based small scale purification of up to 96 proteins using a semi-automated robotics) are all available to enhance productivity. Advanced, modern liquid chromatography equipment (Akta Pure) provides sophisticated unattended multi-column technology to optimise the subsequent protein purification steps.

Biophysical analysis provides scientists access to a wide range of techniques for measuring various protein-protein, protein-ligand interactions, affinities and kinetics. These include Microscale Thermophoresis (MST), Bio-Layer Interferometry (Octet), size exclusion chromatography coupled with multi-angle static laser light scattering (SEC-MALLS), Isothermal Titration Calorimetry (ITC), Dynamic Light Scattering (DLS), Circular Dichroism (CD), Analytical Ultracentrifugation (AUC), stopped/quenched flow kinetics and fluorescence (steady-state intensity, polarization, and fluorescence resonance energy transfer).

Protein crystallisation and structure solution provides valuable information about biological systems under study and is supported within the STP by a range of advanced automation solutions. The platform has sophisticated pipetting and imaging robotics to enable the screening of hundreds of crystallisation conditions. With the help of the team, initial crystal hits can be assessed and optimised further with the ultimate goal of achieving diffracting protein crystals. State-of-the-art in-house X-ray equipment allows screening of suitable crystals for subsequent analysis at the Diamond Light Source, UK, and the European Synchrotron Research Facility, France. The dedicated high-end computing within the platform will be used to determine the 3D model of the molecule under investigation.

The SB STP works extensively with programme leaders who specialise in structural biology within the Francis Crick Institute and who will benefit from access to well managed facilities. We also interact with many programme leaders who are not specialists in structural biology with the objective of adding scientific value to their research programs, encouraging new interactions with scientists who might otherwise have not considered structural biology. Engaging with the SB STP will reduce the barriers associated with picking up new research methods making it possible for groups to envisage novel directions for their research.

Philip Walker Structural Biology lead portrait

Phil Walker (Head) 

philip.walker@crick.ac.uk
+44 (0)20 379 61670

Svend Kjaer (Deputy Head) 

svend.kjaer@crick.ac.uk
+44 (0)20 379 61699