- Total grants
- Total funders
- Total recipients
- Earliest award date
- 10 Apr 2001
- Latest award date
- 30 Sep 2018
- Total GBP grants
- Total GBP awarded
- Largest GBP award
- Smallest GBP award
- Total Non-GBP grants
A large number of human diseases are associated with the deposition of amyloid-like aggregates. As amyloid species associated with different diseasesshare structural properties, it is hypothesised a generic mechanism of cytotoxicity may be the underlying cause for many different diseases pathology. The projects key goals are; 1) Elucidate structural information on amyloid fibril ends Ends of fibrils have been shown to interact with synthetic membranes, and so elucidating the structure of fibril ends may provide insights into mechanism of interaction, which may be a pathway of fibril cytotoxicity. 2) Analyse how amyloid fibrils interact with membrane fractions of cells As fibrils have been shown to interact with synthetic membranes, we seek to analyse how they interact with membranes extracted from the cell such as lysosomal membranes. 3) Assess the structure of amyloid in the cellular environment and characterise the cell's response. We aim to image both fibrils and oligomers inside whole cells using high resolution imaging techniques including cryo-electron tomography, in order to characterise how the cell responds to amyloid exposure. These goals will help to elucidate underlying mechanisms of amyloid cytotoxicity, providing insights into the pathology of diseases such as Alzheimer's disease, Parkinson's disease and haemodialysis related amyloidosis.
This proposal aims to identify new genes and proteins involved in eye development by studying a cohort of consanguineous families with recessively inherited microcornea, cataract and iris coloboma. This combination of signs and inheritance pattern has not been reported before, though similar conditions have been described. Preliminary analysis showed that two of these families map to new genetic loci on chromosomes 10 and 20 while a third has a mutation in a gene only previously implicated in ca taract alone. I now propose to complete the genetic analysis of the remaining families, screen known genes where they are implicated by the genetics and look for mutations in new eye development genes at the new loci. Initially I will prioritise positional candidate genes then sequence them directly, but if necessary I will use Next Generation sequencing to generate a complete list of all sequence variants within one of the loci to ensure I find the gene involved. Once I have found such a gene I will use the techniques of molecular and cellular biology to characterise it further in order to better understand how the eye develops and how mutations in this gene cause an inherited developmental defect.
Molecular motors such as myosin operate under conditions where they are pulling against a load, yet they have only been imaged bound to actin under conditions where there is no load. We wish to visualize the effect of strain on the conformation of myosin-5 motor and lever arm domains and to understand how strain affects the myosin-5 kinetics at a single-molecule level. We also wish to study the elasticity and other structural properties of myosin-5 tail which presumably enables myosin to move cargo through meshworks of actin. We have devised a method for answering these questions using single-molecule biophysical techniques.
The goal of this proposal is to gain a mechanistic understanding of how cytoplasmic dynein, the large microtubule-based motor protein, is regulated to produce essential movement of cargo towards the cell interior (towards the microtubule minus-end). An initial step in many dynein transport events is its targeting to the plus-ends of microtubules, which grow and shrink near the cell periphery. Here, dynein is thought to search-and-capture cargos before transporting them towards the minus-end. T he recent advance of a dynein expression system in yeast makes in vitro reconstitution and detailed dissection of these critical regulatory processes possible. I will establish the minimal proteins required to target dynein to the microtubule plus-end, and analyse their behaviour using single-molecule fluorescence microscopy. The structural basis for control of dynein s motility will be elucidated by single-particle electron microscopy and chemical crosslinking, using non-natural amino acids to precisely insert reactive probes within dynein s large structure. I will also isolate a dynein cargo protein and use optical tweezers to test if it forms a load-bearing interaction with dynein. This combination of biochemical, biophysical and structural approaches will shed new light on the regulation of motor proteins inside living cells, which I will develop into a molecular movie.
Engineering Solutions for an Ageing Population with Musculoskeletal & Cardiovascular Disease. 50 more years after 50. 20 Jul 2015
The ageing population is increasing in number and life expectancy. The population expects fifty more years after fifty with high levels of activity and quality of life. However, the musculoskeletal and cardiovascular systems age and degenerate, adversely affecting mobility, ability to work and quality of life. Advances in engineering and bioscience have created opportunities for novel devices and regenerative therapies, which utilise innovative biomaterials or biological scaffolds to guide the patient's own stem cells to repair degenerative tissues. Advances in patient imaging and diagnostics are enabling earlier disease diagnosis with opportunities to intervene earlier in the degenerative process and preserve healthy tissue, and potential to provide patient specific continuum of care. WELMEC will deliver: - Longer lasting joint replacements in the hip, knee and spine. - Novel regenerative biological scaffolds for degenerative joint tissues, dental reconstructions and cardiovascular surgery. - Advances in cell therapies using the patient's stem cells. - Advanced medical imaging to facilitate earlier diagnosis and intervention. - Novel protein biosensors for disease diagnosis and improved patient targeting. WELMEC will integrate over 200 engineering, physical science, life science and medical researchers with clinicians and industrialists to develop and deliver innovative therapies and patient services for the ageing population.
A major aim of the project includes investigating the early protein-protein interactions involved in the formation of amyloid fibrils from a beta2m variant with a D76N amino acid change, which has been identified as the causative agent in a hereditary, systemic and fatal amyloidosis disease. The D76N associated disease has a different pathology to that of wild-type beta2m associated dialysis related amyloidosis therefore a key goal is to understand how the amino acid change D76N affects the structure and dynamics of the protein and how this results in altered pathology. Another key aim is to refine the structure of the kinetically trapped, inhibitory complex formed between murine beta 2m (mbeta2m) and the truncated form of human beta2m DeltaN6, known to promote beta2m aggregation. A combination of small molecule and peptide screening will be employed to identify an inhibitor of DeltaN6 aggregation, guided by structural information about the DeltaN6-mbeta2m interface.
Early Events in the JC Polyomavirus Lifecyle 14 Jul 2014
JC Polyomavirus (JCPyV) causes a rapid demyelinating disease, Progressive Multifocal Leukoencephalopathy (PML), in the immune compromised host. Owing to the increase in AIDS in the developing world and the introduction of immune modulating therapies, JC is an emerging pathogen and there are currently no antiviral therapies that specifically target it. Although there has been much research on JC biology, various aspects of the virus lifecycle are not well understood. In this project, we will gain greater insight into the viral lifecycle by utilizing chemical genetics, biochemical assays and cell biological techniques combined with high resolution imaging. Specifically, we will identify the intracellular route of JC infection from initial receptor engagement to nuclear entry, elucidate host proteins that bind to the virus genome to modulate viral infection and dissect the signalling pathways required for productive JC virus infection. Greater understanding of these aspects of the virus lifecycle may provide novel targets for intervention against PML and underpin the discovery of new therapeutics, whilst increasing our knowledge of JC virus biology.
Determining the mechanism of β-barrel assembly machinery (BAM) in bacterial outer membranes 14 Jul 2014
Outer membrane proteins (OMPs) in Gram negative bacteria are critical for bacterial survival and virulence 1 . However how these beta-barrels fold in the membrane is not well understood. The in vivo folding of most substrate OMPs relies on the function of the beta -barrel assembly complex (BAM)2, of which the key component is BamA 3, itself a beta-barrel. Here we propose to use a panoply of structural and biophysical methods to dissect the functionality of BamA and the mechanism of OMP folding. Our key aims are: 1) To investigate the hypothesis that BamA functions by lateral gating. Using a combination of disulphide linking and FRET we will determine whether lateral gating is necessary for the folding of different OMPs, and how it may function. 2) To determine how BamA function is affected by liposome size and membrane crowding, by utilising a varied subset of lipid types and comparison of substrate OMPs in folding assays. 3) To determine the importance, and role of the beta -signal of OMPs in the interaction with BamA by dynamic force spectroscopy measurements. The question of how OMPs fold represents a fundamental gap in structural biology understanding. In addition, the OMPs of Gram negative bacteria are key to their pathogenicity, therefore understanding the mechanism of BAM may present new possibilities for drug targets.
Polyketides represent a broad and diverse class of natural products which includes many pharmaceutically-relevant compounds. Historically, identification of novel polyketides relied on natural screening methods; however, rational engineering of polyketide synthases (PKS) represents a powerful tool to create libraries of ‘unNatural’ products to be screened for novel therapeutic properties. The project’s key goals are 1) Elucidating the structure of the indanomycin PKS, starting with the first subunit, IdmL By elucidating the structure of indanomycin PKS, we will increase our understanding of the interfaces between the functional domains and the rules governing the successful rearrangement of polyketide assembly lines. 2) Engineering mutant and chimeric IdmL for the production of novel polyketides By employing synthetic biology approaches, we will investigate the potential for engineering the indanomycin gene cluster to produce novel polyketides. 3) Characterising the structure and function of the post-PKS cyclase IdmH To expand a synthetic biologist’s toolkit of natural product modifying enzymes, we aim to elucidate the structure and reaction mechanism of this novel enzyme. These goals will increase our knowledge of the assembly of the PKS complexes as well as the chemistry involved in the generation of mature polyketide and bring us closer to rational engineering of these systems.
Signalling from Receptor Tyrosine Kinases under non-stimulated conditions and the impact on cancer aggressiveness 14 Jul 2014
Receptor Tyrosine Kinases (RTKs) relays signals which regulate processes such as cell growth, differentiation and cell division. Perturbations of signalling cascades can have serious consequences for the cell. It has previously been shown that proteins containing an SH3 domain can interact with a proline-rich region at the intracellular part of the RTK Fibroblast Growth Factor Receptor 2 (FGFR2), which consequently either inactivates the receptor or relays a downstream signal without any extracellular stimuli. Preliminary data suggests that FGFR2 and other RTK peptides containing the proline-rich motif can interact directly or indirectly with proteins containing SH3 domains. The aims of this project are to verify the interaction between FGFR2 and the proteins Fyn and LASP1, and also the interaction between the RTK ERBB2 (Epidermal Growth Factor family member) and Fyn, and possibly LASP1. Further investigation of the signalling pathways under non-stimulated conditions will be carried out, and their implications in cancer metastasis.
Apoptosis is a form of programmed cell death, the avoidance of which is a hallmark of all forms of cancer. The ability to reactivate apoptosis in tumourcells therefore constitutes a rational strategy for the development of effective cancer treatments. Apoptosis is predominantly initiated in cells viathe intrinsic pathway, which is triggered by various cellular stresses and culminates in cell death following cleavage of many cellular components. This pathway is modulated by the Bcl-2 protein family, whose pro- and anti-apoptotic members react to intracellular signals and interact to determine cellular fate. If pro-apoptotic signals predominate, a Bcl-2 family member called Bax moves from the cytosol to accumulate at the mitochondria, where it causes the mitochondrial membrane to permeabilise, releasing cytotoxic molecules into the cytosol. This constitutes the point of no return for the triggering of cell death. This project will identify the various protein partners that bind to Bax during its transformative journey from quiescent cytosolic molecule to cellular executioner at the mitochondrial membrane. It will aim to characterise the binding interactions between Bax and its partners, with the goal of elucidating mechanisms of Bax activation that can be used to inform the development of novel cancer therapies.
Using Cholera toxin B-chain as a system for the targeted delivery of proteins to motor neurons 15 Jul 2013
The aims of the project are to design a method for the reversible binding of antibody-like proteins to Cholera Toxin B subunit (CTB) to facilitate their trafficking into cells. Initially, a screen against CTB will be carried out toidentify suitable Adhirons. These will be conjugated to a range of different proteins, before being combined with CTB, and administered to assess its ability to deliver the complex into motor neurons in vivo. There is currently no method to deliver antibody-like molecules into motor neurons selectively. Sublingual injection of CTB allows this group of neurons to be targeted specifically. This project therefore focuses on the design of an enabling technology to permit delivery of proteins into specific cell typesto address questions in neurobiology. By combining techniques from molecular biology and chemistry to generate the binding complex, and assessing the effect of these complexes on the nervous system in vivo, this project will provide an interdisciplinary approach to understanding neuronal function at the molecular level. Therefore, this project addresses one of the Wellcome Trusts five major research challenges, i.e., understanding the brain.
Molecular basis of biological mechanisms. 15 Jul 2013
BK polyomavirus is the etiological agent in a number of diseases in the immunosuppressed such as: polyomavirus associated nephropathy in kidney transplant patients, haemorrhagic cystitis in bone marrow recipients and HIV-associated salivary gland disease. Despite this, there are currently no effective anti-viral agents targeting this virus. High-resolution structural information on the native virion and the conformational changes it undergoes during host cell infection are currently lacking. Using state of the art cryo-electron microscopy techniques I will achieve the following goals: 1) Determine the structure of native BKPyV capsid and packaged dsDNA genome 2) Characterise structural changes within the capsid of BKPyV during host cell infection 3) Determine the asymmetric structure of the minor capsid proteins VP2 and VP3 4) Use structure based drug design to identify inhibitors of cell entry and structurally characterise their mode of binding By successfully achieving these research objectives, we will gain significant insights into the various structural configurations adopted by this clinically relevant pathogen during host cell infection and in doing so create a platform for structure based drug design of novel antivirals.