a state of the art facility for the study of protein trafficking in vivo
Lead Research Organisation:
University of Leeds
Department Name: Institute of Membrane & Systems Biology
Abstract
Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.
Technical Summary
Determining protein localisation and dynamics is important for answering many questions in biology. To understand how proteins function and are regulated in vivo, we need approaches by which we can determine where proteins go and when, as well as when and where two proteins interact. New and emerging technologies will go a long way towards helping us answer these questions. The first is the development of photo-activatable GFP (PA-GFP). By tagging proteins with PA-GFP, and then using photo-activation to observe a subset of fluorescently labelled molecules on a low fluorescent background, their fate can be accurately determined. The second is the recent developments in the GFP and RFP fluorophores that have improved behaviour in FRET, enabling us to use this approach to investigate protein-protein interactions in vivo. Furthermore, microscopy is now being developed as a tool for high throughput screening approaches, to investigate the effects of mutations, or for screening large numbers of small molecules for ones that have useful effects in cell biology. The major goal of this application is to upgrade our existing bio-imaging facility into a state-of-the-art facility that can exploit these new technologies, with the focus of studying protein trafficking in vivo.
Publications
Whibley C
(2010)
Wild-type and Hupki (human p53 knock-in) murine embryonic fibroblasts: p53/ARF pathway disruption in spontaneous escape from senescence.
in The Journal of biological chemistry
Smith AJ
(2010)
Voltage-dependent charge movement associated with activation of the CLC-5 2Cl-/1H+ exchanger.
in FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Emmott E
(2008)
Viral nucleolar localisation signals determine dynamic trafficking within the nucleolus.
in Virology
Bottanelli F
(2011)
Vacuolar transport in tobacco leaf epidermis cells involves a single route for soluble cargo and multiple routes for membrane cargo.
in The Plant cell
Hall K
(2010)
Unity and diversity in the human adenoviruses: exploiting alternative entry pathways for gene therapy.
in The Biochemical journal
Xu SZ
(2008)
TRPC channel activation by extracellular thioredoxin.
in Nature
Rose K
(2011)
Transcriptional repression of the M channel subunit Kv7.2 in chronic nerve injury.
in Pain
Bubeck J
(2008)
The syntaxins SYP31 and SYP81 control ER-Golgi trafficking in the plant secretory pathway.
in Traffic (Copenhagen, Denmark)
James NJ
(2010)
The role of Cajal bodies in the expression of late phase adenovirus proteins.
in Virology
Prandovszky E
(2011)
The neurotropic parasite Toxoplasma gondii increases dopamine metabolism.
in PloS one
Hong Y
(2012)
The impact of amyloid precursor protein signalling and histone deacetylase inhibition on neprilysin expression in human prostate cells.
in International journal of cancer
Mankouri J
(2008)
The hepatitis C virus non-structural protein NS5A alters the trafficking profile of the epidermal growth factor receptor.
in Traffic (Copenhagen, Denmark)
Nyathi Y
(2010)
The Arabidopsis peroxisomal ABC transporter, comatose, complements the Saccharomyces cerevisiae pxa1 pxa2Delta mutant for metabolism of long-chain fatty acids and exhibits fatty acyl-CoA-stimulated ATPase activity.
in The Journal of biological chemistry
DaSilva LL
(2006)
Targeting of the plant vacuolar sorting receptor BP80 is dependent on multiple sorting signals in the cytosolic tail.
in The Plant cell
Musa H
(2006)
Targeted homozygous deletion of M-band titin in cardiomyocytes prevents sarcomere formation.
in Journal of cell science
Street M
(2006)
Stimulation of Galphaq-coupled M1 muscarinic receptor causes reversible spectrin redistribution mediated by PLC, PKC and ROCK.
in Journal of cell science
Taneja TK
(2009)
Sar1-GTPase-dependent ER exit of KATP channels revealed by a mutation causing congenital hyperinsulinism.
in Human molecular genetics
Bradley H
(2011)
Residues 155 and 348 Contribute to the Determination of P2X7 Receptor Function via Distinct Mechanisms Revealed by Single-nucleotide Polymorphisms
in Journal of Biological Chemistry
Emmott E
(2010)
Quantitative proteomics using SILAC coupled to LC-MS/MS reveals changes in the nucleolar proteome in influenza A virus-infected cells.
in Journal of proteome research
McHale R
(2010)
Prussian blue coordination polymer nanobox synthesis using miniemulsion periphery polymerization (MEPP).
in Chemical communications (Cambridge, England)
Naylor J
(2010)
Pregnenolone sulphate- and cholesterol-regulated TRPM3 channels coupled to vascular smooth muscle secretion and contraction.
in Circulation research
Smith A
(2009)
Potassium Channels
Description | The aim of this project was to improve our ability to use light microscopy to image cells, and within cells. The funding allowed us to buy additional equipment to upgrade our existing confocal microscopes, so that we could improve our imaging. These microscopes are used by over 20 different research groups within the Faculty of Biological Sciences at the University, and have supported a wide range of research, from imaging organelles and how they move in living plants, to imaging receptors in mammalian cells. |
Exploitation Route | The research can be used by those interested in developing treatments for infections and disease (e.g. pharma companies, clinicians). Imaging is central to understanding the healthy human organism, plants and animals. Without knowledge of how things work, it is very difficult to understand what goes wrong in disease states. The new microscopes are essential for using imaging to understand cellular processes, and in detecting what goes wrong in diseases from virus infections, to inherited mutant |
Sectors | Healthcare Pharmaceuticals and Medical Biotechnology |
URL | http://www.fbs.leeds.ac.uk/facilities/bioimaging/ |
Description | This funding provided an upgrade to our bio-imaging facility which is used by over 40 different research groups across biological sciences and medicine. It has had impact in a broad range of healthcare and biological sciences. |
First Year Of Impact | 2007 |
Sector | Healthcare,Other |
Impact Types | Economic |