E bapala karolo ea bohlokoa lits'ebetsong tsa bophelo, phosphorylating Tyrr, Ser, Thr, e fumanehang ho li-peptide.Phosphorylated amino acid e atisa ho sebelisoa tlhahisong ea phosphoric acid peptide, e sebelisoang hona joale ho single phenyl phosphorylated amino acid.HBTU/phosphorylated amino acid hangata e sebelisoa ho hokahanya mokhoa oa HOBt/DIEA oa phosphorylated amino acid, empa phosphoric acid peptide e khetha bofokoli ba mokhoa ona, haholo-holo ho peptide e hlahisoang nako e telele haholo kapa amino acid phosphate, e hokahanyang ts'ebetso e tlase, e tlase. bohloeki ba sehlahisoa sa ho qetela.Bakeng sa phosphopeptide ena, re ile ra nahana ka mokhoa o ikhethileng oa post-phosphorylation.Ts'ebetso ea ho hlahisa ke ho tlosoa ka mokhoa o ikhethileng ha lihlopha tse sireletsang mahlakoreng a amino acid tse hlokang ho ngoloa ka mor'a peptide synthesis.Bakeng sa Tyr, Thr e ka itšoara ka ho toba ka ho sebelisa li-amino acid tse nang le liketane tse sa sireletsoeng.-Ser (trt), e tlosoa ka bongata sebakeng sa 1% TFA / DCM.
Phosphorylation, e nang le double benzene phosphite phosphoramide le naetrojene e mene e hlahisoa 'me phosphoramide tetrazolium e sebetsang lipakeng, e hokahane le hydroxy, ebe tlas'a sebopeho sa acid peroxide oxidation phosphoryl, karabelo e felletseng.
Hajoale, mokhoa oa phetoho oa peptide phosphorylated ho na le mefuta e 'meli ea mantlha:
(1) kenyelletso e tobileng ea li-amino acid tse nang le phosphorylated ka tatellano ea polypeptide;
(2) ka mor'a hore resin e hlahisitsoeng ka tatellano ea peptide, ho Ser, Tyr, kapa Thr phosphorylation ea side chain hydroxyl.
1) e tla netefatsa hore phosphorylated amino acid e tla hlahisa tatellano ea peptide ka kotloloho:
Ke hore, li-amino acid tse hlokang ho phosphorylated li tla ba phosphorylated le ho sireletsoa ka nepo esale pele ho sebelisoa (Thr,Ser kapa Tyr), ebe phosphorylation e tla fokotsoa ho fihlela sebakeng sa khetho ea index ea polypeptide ho latela ts'ebetso e tloaelehileng ea tlhahiso ea SPPS. .Mokhoa ona o bonolo ho sebetsa, e se e le mokhoa o ka sehloohong oa phetoho ea phosphorylated likhoebong tsa peptide.
2) kamora hore resin e hlahisoe ka tatellano ea peptide, ho Ser hydroxy phosphorylation, Tyr, kapa Thr side liketane:
Mokhabiso oa phosphorylation o sebelisa phosphorylation ea konteraka e le 'ngoe ho mokhoa oa polypeptide, ka lebaka la tšitiso e matla e hlahisoang ke lihlopha tse kholo mokhabisong oa ketane ea mahlakoreng, ho thata hore phosphorylation ea li-amino acid e honyehe le ketane ea peptide.Nakong e tlang, ho kenngoa ha li-amino acid ho tla ba thata le ho feta, haholo-holo mokhabiso o nang le libaka tse ngata tsa phosphorylation, moloko o tla ba thata haholo, sebopeho sa sehlahisoa sa ho qetela se rarahane, ho thata ho arohana, 'me chai e tlaase haholo.
Ka hona, ha libaka tse ngata tsa ketane ea peptide li le phosphorylated, motho a ka khetha ho hlahisa tatellano ea peptide holim'a resin ebe e etsa phosphorylate ka sehlopha sa ketane ea hydroxyl ea Ser, Tyr, kapa Thr: moloko oa eona o kenyelletsa ho tlosoa ka mokhoa o ikhethileng. sehlopha sa ts'ireletso sa ketane ea mahlakoreng se hlokang ho amoheloa ha li-amino acid ka mor'a hore peptide e kopane.Bakeng sa Tyr, Thr e ka sebelisa ka ho toba liketane tsa mahlakoreng a amino acid ha li sirelelitsoe karabelong.
Setsi sa ts'ireletso ea ketane ea mahlakoreng 1% TFA / DCM tikoloho e ka tlosoa ka bongata.Ha mokhoa ona o khethoa, phosphoramidite tetrazolium e sebetsang lipakeng e ka hlahisoa ho sebelisoa dibenzyl phosphoramidite le tetrazolium, e amanang le sehlopha sa hydroxyl, ebe oxidized ho theha sehlopha sa phosphoryl tikolohong ea peroxyacid ho phethahatsa karabelo.
Nako ea poso: Sep-08-2023