Ọ na-arụ ọrụ dị mkpa na usoro ndụ, phosphorylating Tyrr, Ser, Thr, nke dị na peptides.A na-ejikarị amino acid phosphorylated eme ihe na mmepụta nke phosphoric acid peptide, nke a na-eji ugbu a na otu phenyl phosphorylated amino acids.HBTU/phosphorylated amino asịd na-ejikarị jikọọ phosphorylated amino asịd HOBt/DIEA usoro, ma phosphoric acid peptide na-ahọrọ nke a ụzọ ntụpọ, karịsịa na emepụtara ogologo peptide peptides ma ọ bụ amino acid phosphate, ijikọ arụmọrụ dị ala, dị ala. ịdị ọcha nke ngwaahịa ikpeazụ.Maka phosphopeptide a, anyị tụlere usoro nhọrọ post-phosphorylation.Usoro ọgbọ bụ ịwepụ nke amino acid dị iche iche na-echebe akụkụ-agbụ nke dị mkpa ka a kpọọ aha mgbe njikọ peptide gasịrị.Maka Tyr, Thr nwere ike meghachi omume ozugbo site na iji amino acid nwere agbụ akụkụ na-enweghị nchebe.-Ser (trt), wepụrụ nke ọma na gburugburu 1% TFA / DCM.
A na-emepụta phosphorylation, nke nwere benzene phosphite phosphoramide okpukpu abụọ na nitrogen anọ na phosphoramide tetrazolium na-arụ ọrụ intermediates, jikọtara na hydroxy, mgbe ahụ n'okpuru acid peroxide oxidation phosphoramide, mmeghachi omume zuru oke.
Ka ọ dị ugbu a, usoro mgbanwe peptide phosphorylated nwere ụdị isi abụọ:
(1) iwebata amino asịd phosphorylated anabataghị nke ọma n'ime usoro polypeptide;
(2) mgbe resin emepụtara na usoro peptide, nye Ser, Tyr, ma ọ bụ Thr phosphorylation nke n'akụkụ yinye hydroxyl.
1) ga-ekwe nkwa phosphorylated amino acid na-ewebata usoro peptide ozugbo:
Ya bụ, amino asịd ndị dị mkpa ka phosphorylated ga-abụ phosphorylated ma chebe ya nke ọma tupu iji (Thr, Ser ma ọ bụ Tyr), mgbe ahụ, a ga-ebelata phosphorylation ahụ ruo n'ókè nhọrọ nke index polypeptide dabere na usoro ọgbọ SPPS nkịtị. .Usoro a dị mfe iji rụọ ọrụ, abụrụla ụzọ bụ isi nke ụlọ ọrụ peptide phosphorylated mgbanwe.
2) mgbe resin emepụtara na peptide usoro, na Ser hydroxy phosphorylation, Tyr, ma ọ bụ Thr n'akụkụ ụdọ:
Ihe ịchọ mma phosphorylation site na iji phosphorylation nke otu nkwekọrịta n'ime usoro polypeptide, n'ihi mgbochi siri ike nke ndị otu buru ibu na-eme n'akụkụ ihe ịchọ mma n'akụkụ, ọ na-esiri ike phosphorylation nke amino asịd na-adaba na yinye peptide.N'ọdịnihu, iwebata amino acid ga-esiwanye ike karị, karịsịa na ihe ịchọ mma nwere ọtụtụ saịtị phosphorylation, ọgbọ ahụ ga-esiwanye ike, ihe mejupụtara ngwaahịa ikpeazụ dị mgbagwoju anya, siri ike ikewapụ, na mkpụrụ dị oke ala.
Ya mere, mgbe ọtụtụ saịtị na peptide yinye na-phosphorylated, onye nwere ike họrọ ịmepụta peptide usoro na resin wee phosphorylate ya n'akụkụ yinye hydroxyl otu Ser, Tyr, ma ọ bụ Thr: ya ọgbọ tumadi na-agụnye nhọrọ nke mwepụ nke. Otu nchebe nke agbụ akụkụ nke chọrọ nnabata nke amino acid mgbe njikọ peptide gasịrị.Maka Tyr, Thr nwere ike iji eriri akụkụ amino acid anaghị echekwa ya na nzaghachi.
Ebe nchebe yinye akụkụ 1% gburugburu TFA/DCM nwere ike wepu ya nke ọma.Mgbe a na-ahọrọ usoro a, enwere ike ịmepụta phosphoramidite tetrazolium na-arụ ọrụ etiti site na iji dibenzyl phosphoramite na tetrazolium, jikọtara ya na otu hydroxyl, wee oxidized na-emepụta otu phosphoryl na gburugburu peroxyacid iji mezue mmeghachi omume ahụ.
Oge nzipu: Sep-08-2023