Ibicuruzwa

Membrane topologiya ya colicin E1 umuyoboro wakozwe na fluorescence resonance ihererekanyabubasha (FRET).FRET yarimo aside irike ya fluorescent amino (coumarin) nkumuterankunga hamwe nibisigisigi byanditseho ibisigazwa bya sisitemu byahujwe na DABMI (4- (dimethylamino) phenylazophenyl-4'-maleimide) nkuwakira FRET.Ibisigazwa bya fluorescent coumarin byinjijwe muri poroteyine binyuze muri orthogonal tRNA / aminoacyl-tRNA synthetase byombi byemerera kwinjiza kurubuga urwo arirwo rwose mumurongo wa colicin.Buri variant yahinduye ihindagurika (TAG) ihinduka rya coumarin hamwe na sisitemu (TGT) ihinduka rya DABMI.Intera esheshatu zifatanije muri helices 1-6 zagenwe hakoreshejwe isesengura rya FRET kubishobora gukemuka hamwe na leta ihujwe na membrane.Amakuru ya FRET yerekanaga impinduka nini mumwanya uhuza intera hagati ya helices 3-6 kumashyirahamwe ya membrane atanga ubushishozi bushya kumiterere ihuza imiyoboro ya domaine.Muri rusange, coumarin-DABMI FRET imikorere yimikorere yagabanutse nyuma yo guhuza membrane, yubakiye kumiterere yumutaka kumuyoboro wa colicin.Icyitegererezo cyigihe cyo gufunga imiyoboro yumuyoboro cyateguwe hashingiwe ku makuru agezweho kandi yatangajwe mbere.Icyitegererezo cyerekana uruziga rwa helices 1-7 mu cyerekezo cyerekezo yisaha uhereye kuruhande rwinyuma zidasanzwe hamwe na membrane interacial association ya helices 1, 6, 7, na 10 hafi yumusatsi wo hagati wa transembrane wakozwe na helices 8 na 9.