Kayayyaki

An yi nazarin ilimin kimiyyar membrane na yankin tashar colicin E1 ta hanyar canja wurin makamashi mai haske (FRET).FRET ta ƙunshi amino acid mai kyalli (coumarin) a matsayin mai ba da gudummawa da zaɓin ragowar cysteine ​​da aka zaɓa tare da DABMI (4- (dimethylamino) phenylazophenyl-4'-maleimide) azaman mai karɓar FRET.Ragowar coumarin mai walƙiya an haɗa shi cikin furotin ta hanyar tRNA/aminoacyl-tRNA synthetase na orthogonal wanda ya ba da izinin haɗawa da zaɓi cikin kowane rukunin yanar gizon da ke cikin yankin tashar colicin.Kowane bambance-bambancen ya ƙunshi maye gurbi tasha (TAG) don haɗawar coumarin da maye gurbin cysteine ​​(TGT) don haɗin DABMI.An ƙaddara nisa guda shida tsakanin helices 1-6 ta amfani da bincike na FRET don duka masu narkewa da jihohin da ke daure.Bayanan FRET sun nuna manyan canje-canje a cikin nisa tsakanin helices 3-6 akan ƙungiyar membrane suna ba da sabon haske game da tsarin da aka ɗaure membrane na yankin tashar.Gabaɗaya, ingantaccen aikin coumarin-DABMI FRET na tsaka-tsakin ya ragu akan ɗaure membrane, yana ginawa akan ƙirar laima don tashar colicin.An ƙirƙira ƙirar ƙira don rufaffiyar yanayin yankin tashar bisa ga bayanan FRET na yanzu da aka buga a baya.Samfurin yana ba da shawarar tsarin madauwari na helices 1-7 a cikin tazarar agogo daga gefen extracellular da membrane interfacial ƙungiyar helices 1, 6, 7, da 10 a kusa da tsakiyar transmembrane hairpin kafa ta helices 8 da 9.