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handout17 - 5–14 EXAMPLE Ryanodine Recepter(RyR[ryrpaper...

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EXAMPLE: Stopped-Flow Method (solution phase) Reactant A Reactant B Mixing Chamber Stopping Syringe Light Detector Reactants are mixed in a chamber Reaction is monitored as a function of distance from the mixing chamber. Can monitor change in absorbance, fluorescence, scattering, circular dichroism (method to detect secondary structure composition of proteins) Resolution up to 1ms, limited by mixing time (diffusion). instruments can handle > 2 reactants. NOTE: Modification: Quench-Flow: stop reaction after a certain time by cooling, adding a inhibitor solution, optical quenching. EXAMPLE: Flash Photolysis IDEA: many reactions need to to be activated. use (laser) light to excite molecules and trigger a reaction. pump pulse: high energy short pulse (as short as femtoseconds) test pulses: to monitor relaxation or progression of reaction
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Unformatted text preview: 5–14 EXAMPLE: Ryanodine Recepter (RyR) [ryrpaper.pdf] PDB entry 2K2F. BACKGROUND: many cellular responses initiated by cytosolic Ca 2 + : (muscle contraction, neurotransmitter release, secretion, gene transcription. .) • release channels: amplify signal by releasing more Ca 2 + • This results in a sudden increase of calcium (’Calcium spark’). NOTE: A number of sicknesses related to incorrect regulation of calcium spark. PROBLEM: membrane bound proteins difficult to study in situ . SOLUTION: Introduce marker molecules whose flourescence depends on calcium concentration. ⇒ can observe calcium spark behaviour in cells in the presence of drugs (e.g. caffeine increases sensitivity ( muscles more twitchy!)) 5–15...
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handout17 - 5–14 EXAMPLE Ryanodine Recepter(RyR[ryrpaper...

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