Tekinoloje ya FRET peptide

Fluorescence resonance energy transfer (FRET)

Fluorescence resonance energy transfer (FRET) ndi njira yosamutsira mphamvu yomwe woperekayo amasangalala nayo imasamutsidwa kupita kudziko losangalala la wolandira kudzera mu mgwirizano wa maanja amagetsi a intermolecular.Izi sizimaphatikizapo ma photon ndipo kotero sizowotcha.Kuyesa uku kuli ndi ubwino wokhala wofulumira, womvera komanso wosavuta.

Utoto womwe umagwiritsidwa ntchito poyesa FRET ukhoza kukhala wofanana.Koma nthawi zambiri amagwiritsidwa ntchito mitundu yosiyanasiyana ya utoto.Mwachidule, kusamutsidwa kwa mphamvu zowunikira zowunikira ndikutengerapo kwa dipoles kuchokera kwa wopereka (dye 1) kupita kwa wolandila (dye 2) pomwe gulu lopereka likusangalala.Nthawi zambiri, mawonekedwe otulutsa a gulu la Donor fluorophore amadutsana ndi mayamwidwe a gulu la Acceptor."Pamene mtunda wapakati pa ma fluorophores awiri uli woyenera (10 - 100 A), kusamutsidwa kwa mphamvu ya fluorophore kuchokera kwa wopereka kupita kwa wolandira kungawonedwe."Njira yosinthira mphamvu zimatengera kapangidwe kake ka receptor:

1. Imasinthidwa kukhala kugwedezeka kwa ma molekyulu, ndiko kuti, kuwala kowala kwa kusamutsa mphamvu kumatha.(The receptor ndi chozimitsira kuwala)

2. Kutulutsa kwake kumakhala kochulukira kuposa cholandirira chomwechi, zomwe zimapangitsa kusintha kwamtundu wachiwiri wa fluorescence spectrum.(Zolandilira ndi zotulutsa zowala).

Gulu la opereka (EDANS) ndi jini yolandirira (DABCYL) amalumikizana mofanana ndi gawo lachilengedwe la HIV protease, ndipo gawolo likapanda kulumikizidwa, DABCYL imatha kuzima EDANS kenako kukhala yosazindikirika ndi fluorine.Pakutha kwa HIV-1 protease, EDANS simazimitsidwanso ndi DABCYL, ndipo EDANS luciferases imatha kuzindikirika.Kupezeka kwa protease inhibitors kungayang'anidwe ndi kusintha kwa mphamvu ya fluorescence ya EDANS.

FRET peptides ndi zida zosavuta kuphunzira peptidase nonspecificity.Popeza machitidwe ake amatha kuyang'aniridwa mosalekeza, amapereka njira yabwino yodziwira zochita za enzyme.Sheen yomwe idapangidwa pambuyo pa hydrolysis ya ma peptide bond ndi wopereka / wolandila imapereka mulingo wa zochitika za enzyme pamagulu a nanomolar.Pamene peptide ya FRET imakhala yosasunthika, imasonyeza kutayika kwadzidzidzi kwa kung'anima kwa mkati, koma pamene chomangira chilichonse cha peptide chotsutsana ndi wopereka / wolandira chikusweka, chimatulutsa kuwala, komwe kungathe kudziwika mosalekeza ndipo ntchito ya enzyme imatha kuwerengedwa.