teknụzụ FRET peptide

Nfefe ume resonance nke fluorescence (FRET)

Fluorescence resonance energy transfer (FRET) bụ usoro nnyefe ike na-abụghị nke ọkụ nke onye nyere onyinye nwere obi ụtọ na-ebufe ike steeti na-anabata ya site na mmekọrịta nke ndị di na nwunye eletriki intermolecular.Usoro a anaghị agụnye photons, ya mere ọ dịghị radiative.Nnwale a nwere uru nke ịdị ngwa ngwa, dị nro ma dị mfe.

Ihe e ji esiji mee ihe na FRET assay nwere ike ịbụ otu.Mana n'ọtụtụ ngwa, a na-eji agba agba dị iche iche n'ezie.Na nkenke, ịnyefe ike resonance na-egbuke egbuke bụ ịnyefe otu ụzọ dipoles site na onye nyere onyinye (ede 1) na onye na-anabata ya (dye 2) mgbe ndị na-enye onyinye nwere obi ụtọ.N'ozuzu, ihe dị iche iche emission nke Donor fluorophore otu na-adaba na absorption spectrum nke otu Acceptor."Mgbe anya dị n'etiti fluorophores abụọ kwesịrị ekwesị (10 - 100 A), enwere ike ịhụ nnyefe nke ike fluorophore site na onye nyere onyinye nye onye na-anabata ya."Usoro nke nnyefe ike dabere na usoro kemịkalụ nke onye na-anabata ya:

1. A na-atụgharị n'ime vibration molecular, ya bụ, ìhè na-egbuke egbuke nke nnyefe ike na-apụ n'anya.(Onye na-anabata ya bụ ọkụ ọkụ)

2. Mgbapụta na-esiwanye ike karịa onye na-anabata ya n'onwe ya, na-ebute redshift na ụdị fluorescence nke abụọ."(Ndị na-anabata ya bụ ndị na-egbuke egbuke).

Ndị na-enye onyinye (EDANS) na ndị na-anabata ya (DABCYL) jikọtara ya na mkpụrụ ndụ ebumpụta ụwa nke nje HIV, na mgbe ewepụghị mkpụrụ ahụ, DABCYL nwere ike mebie EDANS wee bụrụ nke a na-apụghị ịchọpụta na fluorine.Mgbe nje HIV-1 kwụsịrị njikọ protease, EDANS anaghịzi emenyụ ya site na DABCYL, na EDANS luciferases nwere ike mechaa chọpụta.Enwere ike nyochaa ọnụnọ nke ndị na-egbochi protease site na mgbanwe na ike fluorescence nke EDANS.

FRET peptides bụ ngwaọrụ dabara adaba iji mụọ nkọwapụta peptidase.Ebe ọ bụ na enwere ike nyochaa usoro mmeghachi omume ya mgbe niile, ọ na-enye usoro dị mma maka ịchọpụta ọrụ enzyme.Sheen a na-emepụta mgbe hydrolysis nke peptide bonds nke onye nyere onyinye / onye na-anabata ya na-enye ihe nleba anya nke ọrụ enzyme na nchịkọta nanomolar.Mgbe peptide FRET adịghị emebi emebi, ọ na-egosi mwepu na mberede nke ọkụ dị n'ime, mana mgbe njikọ peptide ọ bụla na-emegide onye nyere onyinye / onye nnabata na-agbaji, ọ na-ewepụta ọkụ, nke enwere ike ịchọpụta ya na-aga n'ihu na arụ ọrụ enzyme enwere ike tụọ ya.