Question

Explain, with the aid of a diagram(s) the physical basis and information that can be gained from Fluorescence Resonance Energy Transfer (FRET) experiments (10 marks) Hand written it possible please.

Answer 1

0Explain with the aid a diagram the Physical basis and in formahon thet cen be gained em Fourescence Resonante Energy Transper (FRET) Exreriment Sep FRET Standor FLoursence Resonance Energy Transer forester Resonance Energy Transfer Singletsinglet Resonance Enerey Taner Resonance Enerey Transfer Tt mainly Used to measure cistance e detect molecular interaction in a daiso uned domains ina single Principle This Expeiment based aner en ergy drom a deonar molecule to an 2Cce phor mole cue number distance etween protein & its conormation s ystem to meagure FRET en the distnce derendert l'e tranter excitahon a donar Fi rophoreo a nearby acteptor enerjy dlorophonre in a non-radiative manner throjh long-renje the distance between lourophores then there elipole-dipol interaction lage OcCur no FRET distance bln fiourophore y Amall OcturA L 9 the FRET Emission Excitahon 4-636nm Emission excitaton C4-33 nm nm 476 mm or 480m Protein fluorescent CFP YFP Nellpw CYAN FloUres cent Pro tein Ecided Stte Shorder wavelegh Hiher eneny & vice er s2 9n Pround s1ate Thi proces fioorescence & the mo1-e cule hat shewn his procesS Frorescent moe cule Dr fIuorophores to0-25oonm Low amount Eneny 1 hign 180-350 roundl Jtate

elaxaton (Ps) FRET (ns) ence So So Areptor DOnov y The inverse sixth Power distane dependene Fars ter Reso Enengy transler wa experimentsk Contimed by wilchek Etdictency o1 FRET TryPtophyl pephdes the Percent2nje inaicates FRET, Ectency the that Contribute ho photons TPA given eA E = +RET here De Flo rescence lie hme v denern the preve nre acceptor & T in the absence cceptor ne more decrezpe an The more tRE Toccu in aoous donar luoreseence lijetime FRET Strongly depends en the dutanre bln the doner & acceptor florophorex (Sixh-Powor rela tionship) The eicienty also depends en the donor to acceptor geperahon distance R inverse 6 order law due t dipole di po le Coupti'ng interaction E (eNctenty) with an R.s her R bem the dis tane e between donor accephor pair & a cceptor at whieh PAEF eiceney s coy between dlonor d forster chs tanee is he Ro ALsorphon(as) orescente Cns