Oye ti peptide phosphoric acid

O ṣe ipa pataki ninu awọn ilana igbesi aye, phosphorylating Tyrr, Ser, Thr, ti o wa lori awọn peptides.Awọn amino acid phosphorylated jẹ eyiti a lo nigbagbogbo ni iṣelọpọ ti peptide phosphoric acid, ti a lo lọwọlọwọ ni awọn amino acids phenyl phosphorylated ẹyọkan.HBTU/phosphorylated amino acids ti wa ni commonly lo lati so awọn phosphorylated amino acids HOBt/DIEA ọna, ṣugbọn awọn phosphoric acid peptide ti wa ni yiyan yi ona abawọn, paapa ninu awọn ti ipilẹṣẹ Elo to gun peptide peptides tabi amino acid fosifeti, sisopọ awọn ṣiṣe ti wa ni kekere, kekere. ti nw ti ik ọja.Fun phosphopeptide yii, a gbero ọna yiyan post-phosphorylation yiyan.Ilana iran jẹ yiyọkuro yiyan ti awọn ẹgbẹ aabo ẹgbẹ-pq amino acid ti o nilo lati jẹ aami lẹhin iṣelọpọ peptide.Fun Tyr, Thr le fesi taara nipa lilo amino acids pẹlu awọn ẹwọn ẹgbẹ ti ko ni aabo.-Ser (trt), pipo kuro ni ayika 1% TFA / DCM.

Phosphorylation, pẹlu ė benzene phosphite phosphoramide ati nitrogen mẹrin ti wa ni ipilẹṣẹ ati phosphoramide tetrazolium ti nṣiṣe lọwọ agbedemeji, ti a ti sopọ si awọn hydroxy, lẹhinna labẹ awọn acid peroxide oxidation phosphoryl formation, pipe lenu.

Lọwọlọwọ, ọna iyipada peptide phosphorylated awọn oriṣi akọkọ meji wa:

(1) ifihan taara ti awọn amino acid phosphorylated ti o ni idaniloju niwọntunwọnsi sinu ọkọọkan polypeptide;

(2) lẹhin ti awọn resini ti ipilẹṣẹ lori peptide ọkọọkan, to Ser, Tyr, tabi Thr phosphorylation ti ẹgbẹ pq hydroxyl.

1) yoo ṣe iṣeduro iwọntunwọnsi awọn amino acids phosphorylated taara ṣafihan ọkọọkan peptide:

Iyẹn ni, awọn amino acids ti o nilo lati wa ni phosphorylated yoo jẹ phosphorylated ati aabo daradara ni ilosiwaju lilo (Thr, Ser tabi Tyr), ati lẹhinna phosphorylation yoo dinku si aaye yiyan ti atọka polypeptide ti o da lori ilana iran SPPS deede. .Ọna yii rọrun lati ṣiṣẹ, ti di ọna akọkọ ti awọn ile-iṣẹ peptide phosphorylated iyipada.

2) lẹhin resini ti ipilẹṣẹ lori ọna peptide, si Ser hydroxy phosphorylation, Tyr, tabi Thr ẹgbẹ ẹwọn:

Ohun ọṣọ phosphorylation nipa lilo phosphorylation ti ihamọ kan sinu ọna polypeptide, nitori idiwọ steric ti ipilẹṣẹ nipasẹ awọn ẹgbẹ nla ni ohun ọṣọ ẹwọn ẹgbẹ, o nira fun phosphorylation ti amino acids lati dinku pẹlu pq peptide.Ni ọjọ iwaju, iṣafihan amino acids yoo nira diẹ sii, ni pataki ninu ohun ọṣọ ti o ni awọn aaye phosphorylation pupọ, iran yoo nira pupọ, akopọ ọja ikẹhin jẹ eka, nira lati yapa, ati ikore jẹ kekere pupọ.

Nitorinaa, nigbati awọn aaye pupọ ninu pq peptide jẹ phosphorylated, ọkan le yan lati ṣe ipilẹṣẹ peptide ọkọọkan lori resini ati lẹhinna phosphorylate nipasẹ ẹgbẹ ẹgbẹ hydroxyl ẹgbẹ ti Ser, Tyr, tabi Thr: iran rẹ ni pataki pẹlu yiyọkuro yiyan ti Ẹgbẹ aabo pq ẹgbẹ ti o nilo idanimọ ti amino acids lẹhin iṣelọpọ peptide.Fun Tyr, Thr le lo taara awọn ẹwọn ẹgbẹ amino acid ko ni aabo ni idahun.

Ipilẹ aabo pq ẹgbẹ 1% TFA/DCM ayika le yọkuro ni iwọn.Nigbati a ba yan ọna yii, agbedemeji phosphoramidite tetrazolium ti nṣiṣe lọwọ le ṣe ipilẹṣẹ nipa lilo dibenzyl phosphoramidite ati tetrazolium, ti o sopọ mọ ẹgbẹ hydroxyl, ati lẹhinna oxidized lati ṣẹda ẹgbẹ phosphoryl ni agbegbe peroxyacid lati pari iṣesi naa.