Ngwaahịa

A mụrụ topology akpụkpọ ahụ nke ngalaba ngalaba colicin E1 site na mbufe ike resonance ume (FRET).FRET tinyere mkpụrụ ndụ ihe nketa nke fluorescent amino acid (coumarin) dị ka onye na-enye onyinye na ihe fọdụrụ nke cysteine ​​a na-ahọrọ nke ejiri DABMI (4- (dimethylamino) phenylazophenyl-4'-maleimide) dị ka onye na-anabata FRET.A na-etinye ihe fọdụrụ na fluorescent coumarin na protein site na orthogonal tRNA/aminoacyl-tRNA synthetase ụzọ nke kwere ka ntinye nhọrọ na saịtị ọ bụla dị na ngalaba colicin.Ọdịiche dị iche iche nwere nkwụsi ike (TAG) maka ntinye coumarin yana ngbanwe cysteine ​​(TGT) maka mgbakwunye DABMI.Achọpụtara anya isii interhelical n'ime helices 1-6 site na iji nyocha FRET maka steeti ndị nwere ike soluble na akpụkpọ ahụ.Ihe omuma FRET gosiputara nnukwu mgbanwe n'ime oghere interhelical n'etiti helices 3-6 n'elu njikọ akpụkpọ anụ na-enye nghọta ọhụrụ n'ime usoro akpụkpọ anụ nke ngalaba ọwa.Na mkpokọta, arụmọrụ interhelical coumarin-DABMI FRET belatara n'elu njide akpụkpọ ahụ, na-ewulite n'ụdị nche anwụ maka ọwa colicin.Emepụtara ihe nlere anya maka ọnọdụ mechiri emechi nke ngalaba ọwa dabere na data FRET dị ugbu a na nke ebipụtara na mbụ.Ihe nlereanya ahụ na-atụ aro nhazi okirikiri nke helices 1-7 na ntụgharị elekere site na akụkụ extracellular na akpụkpọ ahụ interfacial njikọ nke helices 1, 6, 7, na 10 gburugburu isi ntutu transmembrane nke etiti hibere site na helices 8 na 9.