Ubuhanga bwa peptide

Fluorescence resonance ihererekanyabubasha (FRET)

Fluorescence resonance ihererekanyabubasha (FRET) nigikorwa cyo guhererekanya ingufu zidafite imirasire aho umuterankunga yishimiye ingufu za leta yimurirwa muri reta yishimye binyuze mumikoranire yabashakanye bafite amashanyarazi.Iyi nzira ntabwo irimo fotone rero ntabwo irasa.Iyi suzuma ifite ibyiza byo kwihuta, kumva kandi byoroshye.

Irangi ryakoreshejwe mubisobanuro bya FRET rirashobora kuba kimwe.Ariko mubisabwa byinshi, amarangi atandukanye arakoreshwa.Muri make, ihererekanyabubasha rya luminous resonance ni ihererekanyabubasha rya dipole kuva kumuterankunga (irangi 1) kubakira (irangi 2) mugihe itsinda ryabaterankunga ryishimiye.Muri rusange, imyuka yoherezwa mu itsinda rya Donor fluorophore iruzuzanya hamwe no kwinjiza ibintu mu itsinda ryakira.Ati: “Iyo intera iri hagati ya fluorofore ikwiye (10 - 100 A), hashobora kugaragara ko ingufu za fluorophore ziva mu baterankunga ku zakira.”Uburyo bwo guhererekanya ingufu biterwa nimiterere yimiti ya reseptor:

1. Yahinduwe mukuzunguruka kwa molekile, ni ukuvuga urumuri rumurika rwo guhererekanya ingufu rurazimira.(Reseptor ni kuzimya urumuri)

2. Ibyuka bihumanya ikirere birakomeye kuruta ibyakiriwe ubwabyo, bikaviramo guhinduka mu cyiciro cya kabiri cya fluorescence. ”(Abakira ni abasohora luminous).

Itsinda ry'abaterankunga (EDANS) na gene yakira (DABCYL) bifitanye isano imwe na substrate naturel ya protease ya virusi itera sida, kandi iyo substrate idahagaritswe, DABCYL irashobora kuzimya EDANS hanyuma igahita itamenyekana kuri fluor.Iyo virusi ya VIH-1 ihagaritswe, EDANS ntikizimye na DABCYL, kandi EDANS luciferase irashobora kumenyekana nyuma.Kuboneka kwa protease inhibitor birashobora gukurikiranwa nimpinduka zuburemere bwa fluorescence ya EDANS.

FRET peptide nibikoresho byoroshye byo kwiga peptidase idafite akamaro.Kubera ko imikorere yacyo ishobora gukurikiranwa ubudahwema, itanga uburyo bworoshye bwo kumenya ibikorwa bya enzyme.Sheen yakozwe nyuma ya hydrolysis yumubumbe wa peptide nuwaterankunga / uwakiriye itanga igipimo cyibikorwa bya enzyme kuri nanomolar.Iyo peptide ya FRET idahwitse, irerekana kubura gutunguranye kwimbere yimbere, ariko mugihe umugozi wose wa peptide uhabanye numuterankunga / uwakiriye wacitse, urekura flash, ishobora kumenyekana ubudahwema kandi ibikorwa bya enzyme birashobora kugereranywa.