Idlala indima ebalulekile ezinqubweni zokuphila, i-phosphorylating Tyrr, Ser, Thr, etholakala kuma-peptides.I-amino acid eyi-phosphorylated ivamise ukusetshenziswa ekukhiqizeni i-phosphoric acid peptide, okwamanje esetshenziswa kuma-amino acid e-phenyl phosphorylated eyodwa.Ama-amino acid e-HBTU/phosphorylated avame ukusetshenziswa ukuxhuma indlela ye-phosphorylated amino acid HOBt/DIEA, kodwa i-phosphoric acid peptide ikhetha le ndlela yokukhubazeka, ikakhulukazi kuma-peptide ama-peptide amade kakhulu akhiqizwe noma i-amino acid phosphate, ukuxhuma ukusebenza kahle kuphansi, kuphansi. ukuhlanzeka komkhiqizo wokugcina.Ngale phosphopeptide, sicabangele indlela ekhethiwe ye-post-phosphorylation.Inqubo yokukhiqiza iwukususwa okukhethiwe kwamaqembu okuvikela aseceleni e-amino acid adinga ukulebula ngemva kokuhlanganiswa kwe-peptide.Ku-Tyr, i-Thr ingasabela ngokuqondile isebenzisa ama-amino acid ngamaketanga aseceleni angavikelekile.-I-Ser (trt), isuswe ngobuningi endaweni engu-1% TFA/DCM.
I-phosphorylation, ene-double benzene phosphite phosphoramide kanye ne-nitrogen emine iyakhiqizwa futhi i-phosphoramide tetrazolium ephakathi nendawo esebenzayo, ixhunywe ku-hydroxy, bese ngaphansi kokwakheka kwe-asidi ye-oxidation phosphoryl, ukusabela okuphelele.
Njengamanje, indlela yokuguqula i-peptide phosphorylated kunezinhlobo ezimbili eziyinhloko:
(1) ukwethulwa okuqondile kwama-amino acid e-phosphorylated aqinisekisiwe ngokulandelana kwe-polypeptide;
(2) ngemva kwe-resin ekhiqizwe ngokulandelana kwe-peptide, ku-Ser, Tyr, noma i-Thr phosphorylation ye-side chain hydroxyl.
I-1) izoqinisekisa isiqinisekiso esimaphakathi se-phosphorylated amino acids yethula ngokuqondile ukulandelana kwe-peptide:
Okusho ukuthi, ama-amino acid adinga ukwenziwa nge-phosphorylated azobe ephosphorylated futhi avikelwe ngokufanele kusenesikhathi kusetshenziswa (Thr,Ser noma Tyr), bese iphosphorylation izokwehliswa ifike endaweni yokukhetha yenkomba ye-polypeptide ngokusekelwe kunqubo evamile yokukhiqiza i-SPPS. .Le ndlela kulula ukusebenza, isibe indlela eyinhloko amabhizinisi peptide phosphorylated ukuguqulwa.
2) ngemva kwe-resin ekhiqizwe ngokulandelana kwe-peptide, ku-Ser hydroxy phosphorylation, iTyr, noma i-Thr side chain:
Umhlobiso we-phosphorylation usebenzisa i-phosphorylation yokufinyela okukodwa endleleni ye-polypeptide, ngenxa yesithiyo esiqinile esikhiqizwe amaqembu amakhulu emhlobiso weketango elisemaceleni, kuba nzima ukuthi i-phosphorylation yama-amino acid ishwabane neketango le-peptide.Esikhathini esizayo, ukwethulwa kwama-amino acid kuzoba nzima kakhulu, ikakhulukazi emhlobiso oqukethe izindawo eziningi ze-phosphorylation, isizukulwane sizoba nzima kakhulu, ukwakheka komkhiqizo wokugcina kuyinkimbinkimbi, kunzima ukuhlukanisa, futhi isivuno siphansi kakhulu.
Ngakho-ke, lapho amasayithi amaningi ochungechungeni lwe-peptide enziwe i-phosphorylated, umuntu angakhetha ukukhiqiza ukulandelana kwe-peptide ku-resin bese eyiphosphorylate ngeqembu le-hydroxyl eseceleni le-Ser, Tyr, noma i-Thr: isizukulwane sayo sibandakanya ikakhulukazi ukususwa okukhethiwe kwe-resin. iqembu elivikelayo le-side chain elidinga ukuqashelwa kwama-amino acid ngemuva kokuhlanganiswa kwe-peptide.Ku-Tyr, i-Thr ingasebenzisa ngokuqondile amaketanga aseceleni e-amino acid awavikelekile ekuphenduleni.
Isisekelo sokuvikela iketango eliseceleni 1% imvelo ye-TFA/DCM ingasuswa ngokomthamo.Uma le ndlela ikhethiwe, i-phosphoramidite tetrazolium esebenzayo ephakathi ingakhiqizwa kusetshenziswa i-benzyl phosphoramidite ne-tetrazolium, exhunywe eqenjini le-hydroxyl, bese i-oxidized ukuze yakhe iqembu le-phosphoryl endaweni ye-peroxyacid ukuze kuqedelwe ukusabela.
Isikhathi sokuthumela: Sep-08-2023