Nā huahana

Ua aʻo ʻia ka topology membrane o ka colicin E1 channel domain e ka fluorescence resonance energy transfer (FRET).Hoʻopili ka FRET i kahi waikawa amino fluorescent i hoʻopili ʻia (coumarin) ma ke ʻano he mea hāʻawi a me kahi koena cysteine ​​i koho ʻia i hoʻopaʻa ʻia me DABMI (4-(dimethylamino)phenylazophenyl-4'-maleimide) ma ke ʻano he mea ʻae FRET.Ua hoʻokomo ʻia ke koena coumarin fluorescent i loko o ka protein ma o ka orthogonal tRNA/aminoacyl-tRNA synthetase pair i ʻae i ka hoʻohui koho ʻana i kekahi pūnaewele i loko o ka domain channel colicin.Loaʻa i kēlā me kēia ʻano ka hoʻololi ʻana (TAG) no ka hoʻohui ʻana i ka coumarin a me ka hoʻololi ʻana o ka cysteine ​​(TGT) no ka hoʻopili DABMI.Ua hoʻoholo ʻia nā mamao interhelical ʻeono i loko o nā helices 1-6 me ka hoʻohana ʻana i ka loiloi FRET no nā mokuʻāina hoʻoheheʻe a me ka membrane.Ua hōʻike ka ʻikepili FRET i nā loli nui i nā mamao interhelical ma waena o nā helices 3-6 ma luna o ka hui membrane e hāʻawi ana i ka ʻike hou i ka hoʻolālā paʻa-membrane o ka domain channel.Ma keʻano laulā, ua emi ka maikaʻi interhelical coumarin-DABMI FRET ma ka hoʻopaʻa ʻana o ka membrane, e kūkulu ana ma luna o ke kumu hoʻohālike no ke kahawai colicin.Ua hoʻomohala ʻia kahi kumu hoʻohālike no ke kūlana paʻa o ke kahawai ma muli o ka ʻikepili FRET o kēia manawa a i paʻi mua ʻia.Hōʻike ke kumu hoʻohālike i ka hoʻonohonoho pōʻai ʻana o nā helices 1-7 ma ke kuhikuhi ʻana i ka uaki mai ka ʻaoʻao extracellular a me ka hui interfacial membrane o nā helices 1, 6, 7, a me 10 a puni ka lauoho transmembrane waena i hana ʻia e nā helices 8 a me 9.