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“SEE-DRUG ” is a FP7 - RESEARH POTENTIAL project that aims to the upgrade the Structural Biology capacities of UPAT and to the Establishment of a Center of Excellence for Structure-Based Drug Target chracterization efforts in South-Eastern EU region.
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“SEE-STRUCT” is structure-oriented module of SEE-DRUG project and aims to the structural elucidation of proteins, enzymes and other interesting drug-targets. The tools offered for these efforts are a state-of-the-art high field NMR instrument of 700 MHz equipped with a highly sensitive probe and crystallization robot.
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“SEE-PROT” & “SEE-PHARM” are the protein production and the preclinical evaluation of molecules, modules of SEE-DRUG project. The tools offered for these efforts are protein production and characterization facilities an upgraded confocal microscope, an intravital microscope and myographs.
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NMR
700MHz
High sensetivity probe
Studies in solution
Conformation/Dynamics
3D Structure determination, mobility, protein-protein interaction, protein-drug interactions
Confocal
In-cell imaging
Leica Microscope
Tandem Scanner
Additional laser
Sub-cellular ablation in live cells, protein-complexes, protein drug interactions in vivo
Robot
Crystal growth
Automation
Crystalization trials
Fast & efficient
Crystal growth optimization, screen of protein targes in paraller with NRM Spectroscopy
Microscopes
Myographs
intravital Microscope
In vivo
Ex vivo
Drug screening for anti-inflammatory compounds, evaluation of vasoactive compounds

Stuctural Biology
NMR & Xray
2H/13C/15N labeling
Selective labeling
Crystallization
Conformation dynamics, Modeling Structure determination, 3D solutions & crystal models, Bioinformatics
Molecular Biology
Biochemistry
Drug-targets
Biochemical pathways
Protein production
High-yield protein expression, labeling & isolation for structural studies, protein biochemistry
Imaging
In-cell studies
Functional imaging
Light Microscopy
FRET measurements
Studies of biomolecular functions through Advanced Light Microscopy, Sub-cellular ablations
Pharmacology
Inflammation
Vascular Biology
Cell-based assays
Signaling pathways
Monitoring cell processes (migration. proliferation, apoptosis), in vivo & ex vivo assays

Expertise Exchange
Twinnings
Knowledge transfer
Best practices
New applications
Exchange of researchers between UPAT and partnering organizations - EU Centers of excellence
Events
Workshops
Structural Biology
Biochemistry
Imaging
Workshops and scientific meetings will be organized at UPAT, Advanced training course in NMR
Dissemination
Conferences
Biomolecular NMR
Life Sciences
Analysis
Invited talks, Meeting participation & Poster presentations by SEE-DRUG members
Brokerage Events
Info-days
Meet the expers
Links with industry
Regional Authorities
Colloquia with invited speakers from academia or industry and private sector stakeholders

Confocal in cell imaging details

The UPAT’s upgraded Confocal Microscopy Facility, is up to our knowledge unique in Greece for its capacities, properties and applications. This set up will allow the integration of the information gained on drug-target structures following their functional characterization (by the SEE-PROT & SEE-STRUCT modules). Insights from structural determination will be validated in live cells, to allow an understanding of how the biomolecule tested affects basic cellular pathways at the molecular levels. In addition, novel bioactive molecules will be evaluated for their efficacy in modulating protein target function and its signaling pathways.

The upgrade of the confocal microscope facility will be complemented by accompanying facilities, already functional in the Medical School of Patras for culturing mammalian cells including tumorigenic cell line and mouse and human stem cells. In addition the Medical School has a fully equipped animal house for animal studies.

The proposed upgrade of the confocal microscope facility will permit the following methodologies, to be provided to users for the “in cell” characterization of protein function and testing mutated proteins and lead compounds.

  • Cell-based assays for lead compound validation and assessing effects of mutants. Viability, apoptosis, proliferation, differentiation, and senescence assays in cultured mammalian cells, tumor cell lines and neural stem cells.
  • FRET measurements by acceptor photobleaching and sensitized emission for assessing protein-protein and protein-compound interactions and protein modification in live cells. Comparative assessment of wild-type and mutant proteins and the effects of the presence of compounds/peptides.
  • Fluorescence Recovery after Photobleaching (FRAP) experiments for the determination of dynamic protein interactions within cells
  • Sub-cellular ablations coupled to time-lapse microscopy for the analysis of dynamic cellular responses to damage.

This tool complements the existing UPAT instrumentation for assessing protein function and determining protein-protein interactions and modifications in cells and for validation of lead compounds-peptides by in-cell dynamic measurements of protein-protein and protein-compound interactions. The effects on protein function of targeted mutations guided by structural information (SEE-STRUCT) will be assessed in vivo, while bioactive compounds and interacting peptides characterized structurally either by NMR or by X-ray will be tested in cells. Current UPAT staff has extensive experience with Molecular Cell Biology, functional imaging methodologies and cell based assays and will support the running of the facility.

Access to this infrastructure will be provided to a wide variety of Greek labs, both academic and non-academic, in Patras, in Greece and the South-East Region. The Confocal Microscopy Node has been successfully operating in UPAT since 2007 and is in high-demand by a large number of teams in UPAT and Greece (University of Ioannina and Athens).

The facility will be in close contact with Advanced Light Imaging Facilities at the European Level. UPAT teams have a long-standing collaboration with Centers of Excellence in Advance Light Imaging in Europe, including the EMBL in Heidelberg, the Max Planck in Dortmund and the NIMR in London.

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  • 01 Jan 2012 Starting data of SEE-DRUG project

  • 23 Feb 2012 Kick-off meeting

  • 23 Apr2012 External Advisory Board meeting

  • 29 Apr 2013 700MHz NMR installed