Kumvetsetsa kwa phosphoric acid peptide

Zimagwira ntchito yofunika kwambiri m'moyo, phosphorylating Tyrr, Ser, Thr, yomwe ili pa peptides.Phosphorylated amino acid imagwiritsidwa ntchito kwambiri popanga phosphoric acid peptide, yomwe panopo imagwiritsidwa ntchito mu single phenyl phosphorylated amino acid.HBTU/phosphorylated amino acid nthawi zambiri amagwiritsidwa ntchito kulumikiza njira ya phosphorylated amino acid HOBt/DIEA, koma peptide ya phosphoric acid ikusankha zolakwika izi, makamaka pama peptide otalikirapo a peptide kapena amino acid phosphate, kulumikiza magwiridwe antchito ndi otsika, otsika. chiyero cha mankhwala omaliza.Kwa phosphopeptide iyi, tidawona njira yosankha pambuyo pa phosphorylation.Njira yopangira ndikuchotsa mwa kusankha kwa magulu oteteza amino acid omwe amafunikira kulembedwa pambuyo pa kaphatikizidwe ka peptide.Kwa Tyr, Thr imatha kuchitapo kanthu mwachindunji pogwiritsa ntchito ma amino acid okhala ndi unyolo wammbali wosatetezedwa.-Ser (trt), yochotsedwa pamlingo wa 1% TFA / DCM chilengedwe.

Phosphorylation, ndi iwiri benzene phosphite phosphoramide ndi anayi asafe amapangidwa ndi phosphoramide tetrazolium yogwira intermediates, olumikizidwa kwa hydroxy, ndiye pansi pa asidi peroxide makutidwe ndi okosijeni phosphoryl mapangidwe, wathunthu anachita.

Pakadali pano, njira yosinthira peptide phosphorylated pali mitundu iwiri ikuluikulu:

(1) kukhazikitsidwa kwachindunji kwa ma amino acid otsimikizika a phosphorylated mu mndandanda wa polypeptide;

(2) pambuyo pa utomoni wopangidwa pamayendedwe a peptide, kupita ku Ser, Tyr, kapena Thr phosphorylation ya mbali chain hydroxyl.

1) adzatsimikizira zolimbitsa phosphorylated amino zidulo mwachindunji peptide zinayendera:

Ndiko kuti, ma amino acid omwe amafunikira phosphorylated adzakhala phosphorylated ndi kutetezedwa moyenera pasadakhale pogwiritsa ntchito (Thr, Ser kapena Tyr), ndiyeno phosphorylation idzachepetsedwa mpaka malo osankhidwa a polypeptide index potengera njira yachibadwa ya SPPS. .Njirayi ndiyosavuta kugwiritsa ntchito, yakhala njira yayikulu yosinthira mabizinesi a peptide phosphorylated.

2) pambuyo pa utomoni wopangidwa pamndandanda wa peptide, kupita ku Ser hydroxy phosphorylation, Tyr, kapena Thr side chain:

Kukongoletsa kwa phosphorylation pogwiritsa ntchito phosphorylation ya mgwirizano umodzi mu njira ya polypeptide, chifukwa cha chotchinga cholimba chomwe chimapangidwa ndi magulu akuluakulu muzokongoletsa zam'mbali, zimakhala zovuta kuti phosphorylation ya amino acid ichepetse ndi unyolo wa peptide.M'tsogolomu, kuyambika kwa ma amino acid kudzakhala kovuta kwambiri, makamaka mu zokongoletsera zomwe zili ndi malo angapo a phosphorylation, m'badwo udzakhala wovuta kwambiri, mawonekedwe omaliza a mankhwala ndi ovuta, ovuta kupatukana, ndipo zokolola zimakhala zochepa kwambiri.

Chifukwa chake, malo angapo mu unyolo wa peptide ali phosphorylated, munthu amatha kusankha kupanga mndandanda wa peptide pa utomoni ndiyeno phosphorylate ndi gulu lam'mbali la hydroxyl la Ser, Tyr, kapena Thr: m'badwo wake umaphatikizapo kuchotsedwa kosankhidwa kwa utomoni. mbali unyolo zoteteza gulu kuti amafuna kuzindikira amino zidulo pambuyo peptide synthesis.Kwa Tyr, Thr akhoza kugwiritsa ntchito mwachindunji maunyolo amino acid omwe satetezedwa poyankha.

M'mbali mwa unyolo chitetezo maziko 1% TFA/DCM chilengedwe amatha kuchotsedwa mochulukira.Pamene njira imeneyi wasankhidwa, phosphoramidite tetrazolium yogwira wapakatikati akhoza kwaiye ntchito dibenzyl phosphoramidite ndi tetrazolium, zogwirizana ndi gulu hydroxyl, ndiyeno oxidized kupanga phosphoryl gulu mu chilengedwe peroxyacid kumaliza anachita.