Idlala indima ebalulekileyo kwiinkqubo zobomi, i-phosphorylating Tyrr, Ser, Thr, ebekwe kwiipeptides.I-amino acid ephosphorylated iqhele ukusetyenziswa kwimveliso ye-phosphoric acid peptide, okwangoku esetyenziswa kwi-phenyl phosphorylated amino acids enye.I-HBTU/phosphorylated amino acids idla ngokusetyenziswa ukuqhagamshela i-phosphorylated amino acids HOBt/DIEA indlela, kodwa i-phosphoric acid peptide ikhetha le ndlela ineziphene, ngakumbi kwiipeptide zepeptide ezide kakhulu okanye i-amino acid phosphate, ukudibanisa ukusebenza kakuhle kuphantsi, kuphantsi. ukucoceka kwemveliso yokugqibela.Kule phosphopeptide, siye saqwalasela indlela ekhethiweyo ye-post-phosphorylation.Inkqubo yokuvelisa kukususwa okukhethiweyo kwamaqela akhuselayo e-amino acid asecaleni ekufuneka abhalwe emva kokwenziwa kwepeptide.KwiTyr, i-Thr inokusabela ngokuthe ngqo usebenzisa i-amino acids ngamatyathanga asecaleni angakhuselekanga.-I-Ser (trt), isuswe ngokobungakanani kwi-1% TFA / DCM bume.
Iphosphorylation, ene-double benzene phosphite phosphoramide kunye nenitrogen ezine zenziwa kunye ne-phosphoramide tetrazolium intermediates esebenzayo, edityaniswe ne-hydroxy, emva koko phantsi kwe-acid peroxide oxidation phosphoryl ukubunjwa, ukusabela okupheleleyo.
Okwangoku, indlela yokuguqulwa kwe-peptide phosphorylated kukho iindidi ezimbini eziphambili:
(1) ukungeniswa ngokuthe ngqo kwe-amino acids ye-phosphorylated eqinisekisiweyo ngokulandelelana kwe-polypeptide;
(2) emva kweresin eveliswe kulandelelwano lwepeptide, ukuya kwi-Ser, Tyr, okanye i-Thr phosphorylation ye-side chain hydroxyl.
1) uya kumodareyitha isiqinisekiso se-phosphorylated amino acids sazisa ngokuthe ngqo ukulandelelana kwe-peptide:
Oko kukuthi, i-amino acids ezifuna i-phosphorylated ziya kuba ne-phosphorylated kwaye zikhuselwe ngokufanelekileyo kwangaphambili usebenzisa (i-Thr, i-Ser okanye i-Tyr), kwaye ke i-phosphorylation iya kuncitshiswa ukuya kwindawo yokukhetha isalathisi se-polypeptide ngokusekelwe kwinkqubo yokuvelisa ye-SPPS eqhelekileyo. .Le ndlela ilula ukusebenza, ibe yeyona ndlela iphambili yamashishini e-peptide ukuguqulwa kwephosphorylated.
2) emva kweresin eveliswe kulandelelwano lwepeptide, ukuya kwi-Ser hydroxy phosphorylation, iTyr, okanye iThr side chain:
Ukuhlobisa i-phosphorylation usebenzisa i-phosphorylation ye-contraction eyodwa kwindlela ye-polypeptide, ngenxa yesithintelo se-steric esiveliswa ngamaqela amakhulu kumhlobiso wecala lekhonkco, kunzima ukuba i-phosphorylation ye-amino acids iyancipha kunye ne-peptide chain.Kwixesha elizayo, ukuqaliswa kwe-amino acids kuya kuba nzima ngakumbi, ngakumbi kumhlobiso oqukethe iindawo ezininzi ze-phosphorylation, isizukulwana siya kuba nzima kakhulu, ukubunjwa kwemveliso yokugqibela kuyinkimbinkimbi, kunzima ukuhlukana, kwaye isivuno siphantsi kakhulu.
Ke ngoko, xa iisayithi ezininzi kwikhonkco lepeptide zinephosphorylated, umntu unokukhetha ukuvelisa ulandelelwano lwepeptide kwiresin kwaye emva koko iphosphorylate ngecala lekhonkco le-hydroxyl iqela le-Ser, Tyr, okanye iThr: isizukulwana sayo sibandakanya ukususwa okukhethiweyo kwe-resin. iqela elisecaleni lekhonkco elikhuselayo elifuna ukuqondwa kwee-amino acids emva kokwenziwa kwepeptide.Kwi-Tyr, i-Thr inokusebenzisa ngokuthe ngqo i-amino acid emacaleni amatyathanga awakhuselwanga ekuphenduleni.
Isiseko sokukhusela i-side chain 1% TFA / DCM bume inokususwa ngokobungakanani.Xa le ndlela ikhethiweyo, i-phosphoramidite tetrazolium ephakathi esebenzayo inokuveliswa ngokusebenzisa i dibenzyl phosphoramidite kunye ne-tetrazolium, edityaniswe neqela le-hydroxyl, kwaye emva koko i-oxidized ukwenza iqela le-phosphoryl kwindawo ye-peroxyacid ukugqiba ukuphendula.
Ixesha lokuposa: Sep-08-2023