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Probing Protein dynamics on surfaces

Introduction Research Publications & Conference Presentations Equipment & Methods INFORMATION links Equipment link

This research was funded (2003-08) by Science Foundation Ireland as part of a Principal Investigator award (02/IN.1/M231) and in part continued under the National Biophotonics Imaging Platform up to 2011. In 2012 an IRCSET funded PhD student will continue with this research.

PROTEINS & SURFACES AN INTRODUCTION:

Understanding the underlying, fundamental processes that govern the interaction of biological systems with surfaces is key to the development of biocompatible medical devices which lead to successful implantable devices.  This in turn translates to improved healthcare, minimally invasive surgery, healthier and prolonged lives. Studies show that the performance of biomaterials depends on how proteins adsorb on the surface which mediates the interaction of living cells with surfaces [1]. There are different experimental methods for measuring the protein adsorption on surfaces [2], but the majority of techniques do not provide structural information about the adsorbed protein.

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Research:

Our research is focused on developing quantitative fluorescence based methods to measure and analyze the interaction of proteins with different surfaces in real time. Some of the projects which we are undertaking include:

• Protein-Surface Interactions:  Quantifying the amount of protein adsorbing on surfaces & understanding structural changes using advanced fluorescence microscopy.
• Protein denaturation:  We are looking at HSA and BSA (as models) to develop a fluorescence based method to study protein conformational changes in solution.
  
• Protein-Nanoparticle Interactions:  Developing fluorescence based methods to study the interaction of nanoparticles with proteins.
• Protein Sizing:  We have also been involved with researchers in the Department of Microbiology to measure the size of novel proteins using FCS and traditional fluorescence spectroscopy methods. 
• Protein-Protein interactions in living cells:  As part of the Systems Biology Ireland Initiative we collaborate with Dr. H.-P. Nasheuer to measure protein-protein interactions in live cells using Fluorescence Correlation Spectroscopy (FCS).
  

Current researchers:

Przemyslaw Zarski (PhD student, 2012-):  protein-surface interactions measured using TIRF microscopy.

Past senior researchers:

Dr. Denisio Togashi (2004-2011):  protein-surface interactions.
Domhnall O'Shaughnessy (PhD student, protein-nanoparticle interactions).

Other Contributing researchers & students:

2009: Loretta Breslin  and Neil Murphy (MSc project students), Edel Houghton (UREKA summer student)
2008: Amandine Calvet (project student), Valerie Murphy (4Y student).
2007: Muireann O'Loughlin (4Y student), Noemie Marguerite (French Undergraduate), Emmanuelle Bays (Swiss, IAESTE trainee.)
2006: Deirdre McMahon (MSc project student).
2005: Margaret Collins (MSc project student).

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Publications and Conference Presentations:

1. Cell cycle-dependent mobility of Cdc45 in living cells determined by fluorescence correlation spectroscopy.  R. Broderick, S. Ramadurai, K. Toth, D. Togashi, A. G. Ryder, J. Langwoski, and H.P. Nasheuer.  PloS One,  7(4): e35537, ( 2012).
   DOI: 10.1371/journal.pone.0035537. 
2. Assessing protein-surface interactions with a series of multi-labeled BSA using Fluorescence Lifetime Microscopy and Förster Energy Resonance Transfer.  D.M. Togashi and A.G. Ryder, Biophysical Chemistry, 152, 55-64, (2010)
   DOI:  10.1016/j.bpc.2010.07.006
   
3. Monitoring Local unfolding of Bovine Serum Albumin during denaturation using steady-state and time-resolved fluorescence spectroscopy.  D.M. Togashi, A.G. Ryder, and D. O'Shaughnessy, Journal of Fluorescence, 20(2), 441-452, ( 2010). DOI: 10.1007/s10895-009-0566-8
   
4. Quantifying Adsorbed Protein on Surfaces using Confocal Fluorescence Microscopy. D.M. Togashi, A. G. Ryder, and G. Heiss,  Colloids and Surfaces B: Biointerfaces. 72(2), 219-229, ( 2009).
   DOI: 10.1016/j.colsurfb.2009.04.007
5. Investigating trypthopan quenching of fluorescein fluorescence under protolytic equilibrium.  D.M. Togashi, B. Szczupak, A.G. Ryder, A. Calvet, and M. OLoughlin,  Journal of Physical Chemistry A, 113(12), 2757-2767, ( 2009).
   Online at ACS: http://pubs.acs.org/doi/full/10.1021/jp808121y .
6. Trigger factor from the psychrophilic bacterium P. Frigidicola is a monomeric chaperone.  S. Robin, D.M. Togashi, A.G. Ryder, and J.G. Wall, Journal of Bacteriology, 191(4), 1162-1168, ( 2009).
   Online here. DOI: 10.1128/JB.01137-08 
7. A fluorescence analysis of ANS bound to bovine serum albumin: binding properties revisited by using energy transfer.  D.M. Togashi and A.G. Ryder.  Journal of Fluorescence, 18(2), 519-526, ( 2008).  
   Online here. DOI: 10.1007/s10895-007-0294-x.
8. Fluorescence Lifetime Imaging study of a thin protein layer on solid surfaces. D.M. Togashi and A.G. Ryder.   Experimental & Molecular Pathology, 82(2). 135-141, ( 2007).
   DOI: 10.1016/j.yexmp.2007.01.005
9. Mobility and distribution of replication protein A in living cells at single molecule level.  C. Braet, H. Stephan, I. Dobbie, D. Togashi, A.G. Ryder, Z. Foldes-Papp, N. Lowndes, and H.P. Nasheuer.  Experimental & Molecular Pathology, 82(2). 156-162, ( 2007).
   DOI: 10.1016/j.yexmp.2006.12.008
10. Time-resolved fluorescence studies on bovine serum albumin denaturation process.  D.M. Togashi and A.G. Ryder,  Journal of Fluorescence, 16(2), 153-160, ( 2006). 
    DOI:  10.1007/s10895-005-0029-9

Some Presentations:

1. Study of Protein Deposition on Co-Polymers with different wetabilites by Confocal Fluorescence Microscopy, L. Breslin, D.M. Togashi, and A.G. Ryder, 42nd IUPAC Congress, Glasgow, 2-7 Aug. 2009.
2. Probing the Structural Changes of Serum Albumin Protein in Thin Adsorbed Layers on Hydrophilic/Hydrophobic Surfaces using a FLIM-FRET approach.  D.M. Togashi and A.G. Ryder.  International Conference on Trends in Bioanalytical Sciences and Biosensors ( ICTBSB-2009), Dublin, 26-27 January 2009.
   
3. Investigating Intramolecular Fluorescence Quenching in Serum Albumin Protein.  M. O’Loughlin, D.M. Togashi, A.G. Ryder Europtrode IX, Dublin, March 30 - April 2, 2008.
   
4. Confocal Fluorescence lifetime imaging (FLIM): a tool for analysis of structure changes of protein adsorbed onto solid surfaces. D.M. Togashi and A.G. Ryder.   10th Conference on Methods and Applications of Fluorescence: Spectroscopy, Imaging and Probes, Salzburg , Austria , 9-12 Sept., 2007.
   
5. Binding Properties Revisited: a fluorescence analysis of ANS bound to bovine serum albumin.  D.M. Togashi and A.G. Ryder.  10th Conference on Methods and Applications of Fluorescence: Spectroscopy, Imaging and Probes, Salzburg , Austria , 9-12 Sept., 2007. Poster available here.
6. Fluorescence study of Bovine Serum Albumin and Ti and Sn Oxide Nanoparticles Interactions.  D.M. Togashi, A.G. Ryder, D. Mc Mahon, P. Dunne, and J. McManus.  European Conference on Biomedical Optics, Munich, Germany, 17-22 June, 2007. 
7. Evaluating the Structure Changes of Albumin Adsorbed onto Solid Matrix with Different Wettabilities by Fluorescence Lifetime Imaging (FLIM).  D. Togashi and A. Ryder,  7th International ELMI meeting, York, England,  17-20 April, 2007.
8. FCS and Fluorescent proteins: measuring diffusion rates, concentrations and protein-protein interactions.  I. Dobbie, H. Stephan, H.-P. Nasheuer, D. Togashi, A. Ryder, and N. Lowndes.  7th International ELMI meeting, York, England,  17-20 April, 2007.
9. Application of fluorescence lifetime imaging on adsorption of bovine serum albumin on solid surfaces.  D.M. Togashi and A.G. Ryder.  Microscopical Society of Ireland's 30th Annual Symposium, NUI-Galway, 30 Aug.-1 Sept., 2006.
10. Real-time measurement of adsorption of bovine serum albumin on silica glass by confocal fluorescence microscopy. D.M. Togashi and A.G. Ryder.  6th International ELMI meeting, Ofir , Portugal , 30 May-2 June, 2006. 

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METHODS FOR STUDYING PROTEIN DYNAMICS:

This will be updated in the coming months.

We also have a range of other research projects where the equipment could be used for protein-surface interaction measurements, a more detailed list of equipment is given on the Equipment Page.

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INFORMATION Links:

These are some of the sites that we regularly use. I hope to add more links and details in the near future. If there are problems with any of the links let me know.

Technical Journals & Societies:

Surface Research & courses:

Polymer-protein surfaces, Switzerland.
Proteins-Polymers-Interfaces Group, at the Department of Bioengineering at the University of Utah.

Information Resources:

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Equipment Links:

This is a selection of useful web-sites from a range of manufacturers who produce the equipment we use.

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