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Ανάπτυξη στοχαστικού μαθηματικού μοντέλου για την προσομοίωση πειραμάτων ανάκτησης φθορισμού μετά από φωτολεύκανση με σκοπό την μελέτη των κινητικών ιδιοτήτων σημασμένων πρωτεϊνών. [ http://www.api.com/lifescience/dv-QLM.html ]. PRESENTATION OUTLINE. FRAP Introduction Deterministic Modeling

moorejeremy
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  1. Ανάπτυξη στοχαστικού μαθηματικού μοντέλου για την προσομοίωση πειραμάτων ανάκτησης φθορισμού μετά από φωτολεύκανση με σκοπό την μελέτη των κινητικών ιδιοτήτων σημασμένων πρωτεϊνών [http://www.api.com/lifescience/dv-QLM.html]

  2. PRESENTATION OUTLINE • FRAP • Introduction • Deterministic Modeling • Stochastic Modeling • Comparison of the two methods • Analysis of GFP-GR kinetics

  3. FRAP: INTRODUCTION [Nature, 2004] Definition:A live-cell-imaging technique used to study the extensive networks of protein–protein interactions that regulate cellular processes. Technique:A pulse of high-intensity light is used to irreversibly photobleach a population of fluorophores in a target region

  4. FRAP: RECOVERY CURVE Reaction: kon,koff Diffusion: Df [http://www.bio.davidson.edu/Courses/Molbio/FRAPx/FRAP.html]

  5. FRAP: DETERMINISTIC MODELING [Sprague et al, 2004]

  6. FRAP: STOCHASTIC MODELING I Application:nuclear mobility of a GFP-tagged glucocorticoidreceptor (GFP-GR) in nuclei of both normal and ATP- depleted cells.Does it binds to the nuclear matrix (3-dimensional filamentous protein )? [Brown R, 1828] Model:Diffusion is seen as a Brownian random motion (Simulation with Monte Carlo methods) GFP diffuses with no preferred direction due to random collisions with surrounding molecules Brownian motion of DNA-tethered beads[http://www.bio.brandeis.edu/~gelles/movies.html]

  7. FRAP: STOCHASTIC MODELING II [Gillespie D, 1977] Model:Reaction is seen as stochastic biochemical pathway (Simulation with Monte Carlo methods) 1. What is the time that a new reaction will happen? 2. What is the next most probable reaction? 3. What will be the change of reactant species after a reaction has occurred?

  8. FRAP: STOCHASTIC MODELING III

  9. FRAP: DETERMINISTIC VERSUS STOCHASTIC MODELING

  10. FRAP: STUDYING THE NUCLEAR MOBILITY OF A GFP-GR I • Population statistics • Tendency towards reaction equilibrium (entropy!)

  11. FRAP: STUDYING THE NUCLEAR MOBILITY OF A GFP-GR II

  12. FRAP: STUDYING THE NUCLEAR MOBILITY OF A GFP-GR III

  13. FRAP: STUDYING THE NUCLEAR MOBILITY OF A GFP-GR IV

  14. FRAP: STUDYING THE NUCLEAR MOBILITY OF A GFP-GR V

  15. FRAP: WHAT MORE CAN STOCHASTIC SIMULATION OFFER • Firmer physical basis • Richer information • Expensive implementation

  16. ACKNOWLEDGEMENTS • ΙΚΥ • TUGRAZ, AUSTRIA

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