Alaabta

Dusha sare ee xuubabka colicin E1 channel waxaa lagu bartay wareejinta tamarta resonance fluorescence (FRET).FRET waxay ku lug leedahay amino acid fluorescent ah (coumarin) ku-deeqaha iyo hadhaaga cysteine-ka ee la xushay oo lagu xidhay DABMI (4- (dimethylamino) phenylazophenyl-4'-maleimide) sida aqbalaada FRET.Hadhaaga koomarin fluorescent waxaa lagu dhex daray borotiinka iyada oo loo sii marayo lammaanaha orthogonal tRNA/aminoacyl-tRNA synthetase taasoo u oggolaatay in lagu daro goob kasta oo ka tirsan kanaalka colicin.Kala duwanaansho kastaa waxa ay haysay beddelka joogsiga (TAG) ee isku darka coumarin iyo beddelka cysteine ​​(TGT) ee lifaaqa DABMI.Lix fogaansho isdhexgal ah oo ku dhex jira helisyada 1-6 ayaa la go'aamiyay iyadoo la isticmaalayo falanqaynta FRET ee labada dawladood ee milmay iyo xuubabka ku xidhan.Xogta FRET waxay muujisay isbeddelo waaweyn oo ku yimid masaafada u dhaxaysa helices 3-6 marka la eego ururka xuubka oo siinaya aragti cusub qaab dhismeedka xuubabka ee qaybta kanaalka.Guud ahaan, wax ku oolnimada coumarin-DABMI FRET isdhexgalka ayaa hoos u dhacay marka la isku xiro xuubka, iyadoo lagu dhisayo qaabka dallada ee kanaalka colicin.Nashqada ku meel gaadhka ah ee gobolka xidhan ee kanaalka ayaa la sameeyay iyadoo lagu salaynayo xogta FRET hadda iyo hore loo daabacay.Qaabku wuxuu soo jeedinayaa habayn wareeg ah oo helices 1-7 ah oo jihada saacada ah ka soo jeeda dhinaca ka baxsan unugga iyo xuubka interfacial ururka helices 1, 6, 7, iyo 10 agagaarka timaha dhexe ee transmembrane oo ay sameeyeen helices 8 iyo 9.