南京大学-杨荣武生物化学-英文版-第一篇-Protein-GFP.pptVIP

* Green Fluorescent Protein (GFP) * Learning Objectives Structure of GFP Mechanism of action Function in nature Medical Relevance A Cool Protein Give off the green light in the presence of UV light Marker of the twenty-first century Nobel Prize in Chemistry 2008 The molecular structure of the archetypical Aequorea victoria green fluorescent protein (GFP) . The polypeptide chain is shown in cartoon format with the chromophore in stick representation. Origin of GFP Jellyfish, Aequorea victoria in 1962 Modified version of GFP Cat Pig Rabbit http://www.cbc.ca/news/yourview/2007/12/uv_light_makes_fluorescent_fel.html Nobel Prize in 2008 Osamu Shimomura Martin Chalfie Roger Y. Tsien /nobel_prizes/chemistry/laureates/2008/ Aequorea victoria Aequorea victoria photoorgans Florescent Zebra XFPs Structure of GFP * Structure Primary 236 AA residues Secondary * 1 β pleated sheet composed of 11 antiparallel strands (48 %) * 5 α helices (5%) 20 β turns (bends) 2 γ turns 12 β bulges 7 β hairpins Supersecondary β barrel Ligands * Fluorophore (Chromophore) * Fluorophore Functional part of GFP Ser – Tyr - Gly Formed by rapid cyclization between Ser65 and Gly67 followed by a slower rate-limiting oxygenation of the Tyr66 side chain Excitation Spectrum vs. Emission Spectrum Chromophore p-hydroxylbenzylideneimidazolinone Ser65-Tyr66-Gly67 Excitation Spectrum Absorb at maximum at 400nm and 470nm Conjugation Emission Spectrum Shift to 505nm and 540nm Responsible for the color * Action of Fluorophore Transduces blue light to green fluorescent light via energy transfer Blue light Resonant stabilization Green light * * Functions in nature Found in the Pacific Northwest jellyfish Aquorea victoria Works as a spontaneous fluorescent protein which transduces the blue chemiluminescence of the aequorin protein into green fluorescent light Serves as a means of communication Attract prey Means of protection DNA repair * Medical Relevance GFP has been extracted