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Iyo membrane topology yecolicin E1 chiteshi chiteshi chakadzidzwa nefluorescence resonance simba rekufambisa (FRET).Iyo FRET yaisanganisira genetically encoded fluorescent amino acid (coumarin) semupi uye yakasarudzika yakanyorwa cysteine ​​residue yakabatanidzwa neDABMI (4-(dimethylamino) phenylazophenyl-4'-maleimide) semugamuchiri weFRET.Iyo fluorescent coumarin residue yakaiswa muprotein kuburikidza ne orthogonal tRNA/aminoacyl-tRNA synthetase pair iyo yakabvumira kusarudzwa kubatanidzwa mune chero saiti mukati mecolicin chiteshi domain.Imwe neimwe yakasiyana yakabata yekumira (TAG) shanduko yekubatanidzwa kwecoumarin uye cysteine ​​​​(TGT) shanduko yeDABMI yekunamatira.Nhanhatu dzinopindirana madaro mukati mehelice 1-6 dzakatemerwa kushandisa FRET kuongororwa kune ese ari maviri soluble uye membrane-akasungwa nyika.Iyo data yeFRET yakaratidza shanduko huru mumatanho epakati pakati pehelices 3-6 pane membrane kusangana ichipa ruzivo rutsva mune membrane-yakasungwa chimiro chechiteshi chechiteshi.Kazhinji, coumarin-DABMI FRET interhelical efficiencies yakadzikira pane membrane inosunga, ichivaka pamusoro peamburera modhi yecolicin chiteshi.A tentative modhi yenzvimbo yakavharwa yechiteshi chiteshi yakagadziridzwa zvichibva pane yazvino uye yakamboburitswa data yeFRET.Iyo modhi inoratidza denderedzwa kurongeka kwehelice 1-7 munzira yewachi kubva kune extracellular side uye membrane interfacial association yehelices 1, 6, 7, uye 10 yakatenderedza nechepakati transmembrane hairpin inoumbwa nehelices 8 ne9.