The Centre for Molecular and Structural Biochemistry is a multidisciplinary biomolecular research centre based in the Schools of Chemistry (CHE) and Biological Sciences (BIO) at UEA, which brings together scientists working at the interface between biology and chemistry in an environment that enables complementary expertises to be applied to important biological problems.
Launched in the early 1990s, the Centre for Metalloprotein Spectroscopy and Biology provided an umbrella for a range of successful interactions between UEA chemists (in CHE) and biologists (in BIO) aimed at solving a range of significant biological problems. The Centre, which was the forerunner of the many inter-disciplinary research centres that have sprung up in the UK and abroad in recent years, built an international reputation for excellence in research and pioneering innovative techniques. As its name suggests, the focus of the CMSB was metalloproteins - proteins that contain metals, which are essential for many key cellular processes. In particular, the work of the Centre focused on the activation and redox cycling of inorganic substrates involved in bacterial nitrogen, oxygen and sulfur cycles, and on metal ion metabolism and metal-microbe interactions. Click here for a more in-depth information about the history of the Centre.
Though the links between chemists and biologists at UEA remain as strong as ever, the focus of their work has broadened significantly in recent years into areas including protein folding, environmental sensing, pathogen-host interactions and cancer biology. To reflect this change a celebratory meeting headlined by the Nobel Laureate Professor Sir John Walker FRS was held in May 2008 and the Centre was renamed the Centre for Molecular and Structural Biochemistry.
Through multi-million pound investment from BBSRC, the Royal Society and the Wellcome Trust, including a £3.4M JIF award ‘Biophysical Chemistry at UEA’ in 2002 led by one of the CMSB’s founding directors, Professor Andrew Thomson FRS OBE, the Centre boasts a broad and unique range of biophysical facilities. These include the UK's first pulsed multi-frequency EPR spectrometer, 500/600/800 MHz NMR instruments, magnetic circular dichroism, protein film electrochemistry and rapid reaction kinetics, supported by a host of additional biophysical and bioanalytical equipment, all housed within new/refurbished laboratory space.
At the beginning of 2010 Dr Nick Le Brun (CHE) became the new Director of the Centre. On his new role, Dr Le Brun said ‘I am delighted to be leading the Centre into what I hope and intend will be a new era of success. Molecular scientists at the chemistry/biology interface have a key role to play in developing new understanding of complex biological systems and in applying this knowledge to help solve a range of clinical and environmental problems. The CMSB is well placed to make a tremendous contribution to this’.
Click here for more information about the aims, roles and activities of the CMSB
Report for CMSB Project 2012: “The relevance of DNA repair pathways to healthy physiology of human lens cells” – Meghan Betts
DNA damage occurs in cells through a variety of routes, resulting in disruption of normal cellular metabolism that can lead to an accumulation of mutations and cell death. A number of repair pathways are vital in optimising cell survival following DNA damage, with recent evidence implicating a role of Ku proteins in the repair of DNA double strand breaks. The primary aim of this project was to assess whether the load of DNA damage is linked to the biochemical and cellular activities of Ku proteins in the human lens.
Oxidative damage in the lens is important in cataract formation, and here H2O2 treatment has been used in both the FHL124 cells and the lens epithelium to initiate this damage. As Ku80 is a repair protein of the non-homologous end joining (NHEJ) pathway, it is believed that knocking down this protein will cause an inability to repair DNA damage appropriately, which may also lead to an increase in cell death. First, it was important to investigate whether the technique to knockdown Ku80 using siRNA worked as expected. Previous evidence has shown that Ku80 can be knocked down in FHL124 cells, and data obtained within this project confirmed that the siRNA transfections to knockdown Ku80 in the lens epithelium were successful (Figure 1). Further experiments confirmed that siRNA knock down expression of Ku80 leads to an increased level of cell death in FHL124 cells treated with H2O2 (data not shown).
This project has been valuable in showing that reduction of expression of Ku80 leads to an increased amount of cell death in human lens cells. Overall, the work done in this project shows promise for future experiments, particularly if siRNA knockdown of Ku80 can be successful in the whole lens.
Figure 1: Immunocytochemistry confirmed knockdown of Ku80 expression in human lens epithelium. (Left) An antibody against human Ku80 visualised its expression in human lens epithelium that had been transfected with siRNA to knockdown Ku80, or with a “scrambled” siRNA as a negative control. Staining with DAPI confirmed that equivalent numbers of cells were visualised in each sample. (Right) Quantification of immunocytochemistry results using Image J determined that ~81% knockdown of Ku80 expression had occurred after the siRNA transfection compared to the scrambled control. Error bars represent the standard error across 300 cells.
The programme for the CMSB Spring Meeting on Thursday 31st May in UEA's Thomas Paine Study Centre Lecture theatre is shown below.
It was an excellent afternoon of science and an opportunity to hear about the work of some of the more recent arrivals on the Norwich Research Park, as well as a plenary lecture from one of the leading lights in metal homeostasis.
13.30 – 13.40 Welcome and introduction
13.40 – 14.10 Dr Andy Gates (BIO)
‘NasS-NasT; a novel nitrate sensor for nitrogen assimilation in Paracoccus denitrificans’
14.10 – 14.40 Dr Andy Round (PHA)
‘Single molecule studies of heterogeneous biomacromolecules and their complexes’
14.40 – 15.10 Dr Sam Fountain (BIO)
‘Working from the outside: ATP, a key signal in pain and inflammation’
15.10 – 15.40 Coffee (TPSC Foyer)
15.40 – 16.10 Dr Vasily Oganesyan (CHE)
‘Bridging the gap between theoretical modelling and EPR spectroscopy: Application to spin labelled proteins’
16.10 – 16.40 Dr Janneke Balk (JIC/BIO)
‘The role of the Fe-S protein Ind1 in the assembly of mitochondrial complex I’
16.40 – 17.35 Prof Nigel Robinson (University of Durham)
‘How cells help proteins to acquire the correct metals’
17.35 – 17.40 Closing remarks
17.40 – Drinks and nibbles (TPSC Foyer)
Dr Andy Gates
Dr Sam Fountain
The meeting was sponsored by the Biochemical Society.
CMSB Summer Bursary Scheme 2012
Applications are invited for the two 7-week CMSB Summer Bursaries available this year. The aim of the bursary scheme is to promote new collaborations between the CMSB and cell/molecular biologists, leading to novel biochemistry projects based on eukaryotic systems with relevance to human health/well-being.
Details of the Summer Bursary Scheme
· Applications must involve at least two NRP-based research groups, one of which must be part of the CMSB, and must be focussed on a eukaryotic system with clear relevance to human health/well-being (but needn’t involve human macromolecules). Either group can lead the application.
· Applications must include a named researcher who will take up the bursary. The scheme is aimed at students who have just finished their degree programmes (who already have significant experience of laboratory-based work), but others are not excluded (i.e. mid-year undergraduates will also be considered).
· Applications will be judged primarily on the quality and strategic relevance of the science, but the background and suitability of the nominated student will also be taken into account.
· Each bursary project will run for 7 weeks and provides £180 per week for the student.
· A brief report on the outcome of the project will be required within one month of the end date; an edited version will appear on the CMSB website.
Download the application form [DOC]
Eukaryotic biochemistry with relevance to human health/well-being
Within the CMSB, studies of systems derived from eukaryotic organisms with direct relevance to human health and well-being are currently under-represented. This is an area which connects with the NRP’s Food and Health Alliance (FAHA), and research that addresses issues related to human health and well-being is a major area of importance identified by the BBSRC in its Strategic Plan (2010-15) and its Delivery Plan (2011-15). There are clear opportunities on the NRP to develop this area of research, with a significant number of groups in BIO, PHA, MED, the IFR and the Norfolk & Norwich University Hospital working on problems associated with key processes in eukaryotic cells. In 2011, to initiate new interactions between eukaryotic cell biologists and biophysical chemists/biochemists, the CMSB launched a summer internship scheme, to provide two 7-week internships to run over the summer of each year. The intention is to support projects that have the potential to exploit, in the relative short term, the in vitro biophysical technologies/expertise available in the CMSB. It is envisaged that projects supported through the scheme will continue, e.g. through undergraduate/Masters’ research projects, PhD studentship applications etc., leading eventually to major applications to Research Council/major charity organisations.
CMSB Bursaries 2011
The two recipients of the CMSB Bursaries in 2011 were Sophie Bennett (working with Andrew Hemmings and Nathalie Juge) and Lloyd Wahl (working with Tharin Blumenschein, Surinder Soond and Andrew Chantry), and reports on their projects can be found below.
Probing the Molecular Basis for Recognition of the Thomsen-Friedenreich Antigen by Human Galectin-3 using X-ray Crystallography
Circulatory human galectin-3 interacts with cancer-specific glycoconjugates such as the Thomsen-Friedenreich carbohydrate antigen (TF; Galβ1-3GalNAcα1-O-Ser/Thr), promoting cancer cell adhesion and metastasis. We have investigated the molecular basis for this interaction by solving the high resolution X-ray crystal structures of the complexes of galectin-3 carbohydrate recognition domain (CRD) with two forms of the TF antigen. The results will help provide a basis for future structure-based anti-cancer drug design. For a more detailed description of the project [PDF].
NMR studies on the WWP2 ubiquitin ligase WW domains as a step towards the design of novel cancer therapeutics
My project over the summer involved working in both Dr Andrew Chantry’s lab and Dr Tharin Blumenschein’s lab on the purification of segments of the WWP2 protein with a view to analysing their structures using 2D and 3D NMR which would lead on to peptide binding studies. A widely reported paper released earlier in the year by Dr Andrew Chantry and Dr Surinder Soond outlined an important role of the isoforms of WWP2, which were found to differentially bind Smads and modulate TGFβ signalling; working on structural and binding studies in this area represented an opportunity to open the door on to potentially interfering with WWP2 modulation of the TGFβ signalling pathway. To obtain high yields of WWP2, E. coli were transformed with plasmid coding for our protein of interest with a polyhistidine tag; large volumes of media were inoculated with successfully transformed colonies and production of the protein was induced in minimal media to allow Nitrogen-15 labelling. Protein was purified from cell lysates using immobilised metal affinity chromatography, the buffer was exchanged and the protein concentrated. Problems with low yield and protein solubility at the high concentrations required for NMR analysis hindered the progression of the project; troubleshooting these problems was frustrating but a great learning experience all the same, and involved interpreting protocols from research papers published by other groups which had experienced similar problems with other proteins. We progressed as far as early NMR analysis on some parts of the protein but the project has a positive future. Collaborating between a laboratory in the school of biological sciences and a laboratory in the school of chemistry was also a brilliant experience, I learnt that collaborating between two schools not only involved a lot of walking (between the two different labs), but also involved a productive fusion of ideas, approaches and perspectives.
Early Years Researchers’ Colloquium Autumn 2011
The next CMSB Early Years Researchers’ Meeting will take place on Friday Oct 7th at the University of East Anglia's Lecture Theatre 4
The current program involves:
13.30 – 13.40 Welcome and introduction
13.40 – 14.10 Florian Hilbers (Max-Planck Institut für Biophysik, Frankfurt)
‘Hydrogen peroxide decomposition by aa3 cytochrome c oxidase from Paracoccus denitrificans’
14.10 – 14.40 James Tolchard (CHE)
‘Mapping the actin-binding region in the Tarp protein from Chlamydia’
14.40 – 15.10 Rebecca Handley (IFR/CHE)
‘PerR; the role of a metalloregulator in peroxide stress survival in Campylobacter’
15.10 – 15.40 Ellis O’Neil (JIC)
‘Towards the in vitro generation of a starch granule’
15.40 – 16.00 Coffee
16.00 – 16.30 Morgan Bye (CHE)
‘Combining NMR with EPR distances and in silico calculations for a more complete protein-protein interaction model’
16.30 – 17.25 Dr Sarah O’Connor (JIC/CHE)
‘Understanding and engineering alkaloid biosynthesis’
17.25 – 17.30 Closing remarks
17.30 – Drinks and nibbles (BIO Atrium)