Yana taka muhimmiyar rawa a cikin tafiyar matakai na rayuwa, phosphorylating Tyrr, Ser, Thr, dake kan peptides.An fi amfani da amino acid phosphorylated wajen samar da peptide phosphoric acid, a halin yanzu ana amfani da shi a cikin amino acid phenyl phosphorylated guda ɗaya.HBTU/phosphorylated amino acid yawanci ana amfani da su don haɗa hanyar phosphorylated amino acid HOBt/DIEA, amma phosphoric acid peptide suna zabar wannan lahani na hanya, musamman a cikin peptides peptides mafi tsayi ko amino acid phosphate, haɗa ingantaccen aiki yana da ƙasa, ƙasa. tsarki na karshe samfurin.Don wannan phosphopeptide, mun yi la'akari da zaɓaɓɓen hanyar post-phosphorylation.Tsarin tsarawa shine zaɓin cire ƙungiyoyin kariya na gefen sarkar amino acid waɗanda ke buƙatar lakabi bayan haɗin peptide.Ga Tyr, Thr na iya amsawa kai tsaye ta amfani da amino acid tare da sarƙoƙin gefe marasa kariya.-Ser (trt), an cire shi da yawa a yanayin 1% TFA / DCM.
Phosphorylation, tare da biyu benzene phosphite phosphoramide da nitrogen hudu ana haifar da su da kuma phosphoramide tetrazolium masu aiki tsaka-tsaki, an haɗa su da hydroxy, sa'an nan kuma a karkashin acid peroxide oxidation phosphoryl samuwar, cikakken dauki.
A halin yanzu, hanyar gyara peptide phosphorylated akwai manyan nau'ikan guda biyu:
(1) gabatarwa kai tsaye na amino acid phosphorylated masu matsakaicin garanti cikin jerin polypeptide;
(2) bayan guduro da aka samar akan jerin peptide, zuwa Ser, Tyr, ko Thr phosphorylation na gefen sarkar hydroxyl.
1) matsakaiciyar garantin amino acid phosphorylated kai tsaye gabatar da jerin peptide:
Wato, amino acid ɗin da ake buƙatar phosphorylated za su kasance a cikin phosphorylated kuma a kiyaye su yadda ya kamata a gaba ta amfani da (Thr, Ser ko Tyr), sa'an nan kuma za a rage phosphorylation zuwa wurin zaɓi na ma'aunin polypeptide bisa tsarin tsararrun SPPS na yau da kullun. .Wannan hanya yana da sauƙin aiki, ya zama babban hanyar peptide Enterprises phosphorylated gyare-gyare.
2) bayan resin da aka samar akan jerin peptide, zuwa Ser hydroxy phosphorylation, Tyr, ko Thr sassan sassan:
Kayan ado na phosphorylation ta amfani da phosphorylation na haɗin gwiwa guda ɗaya a cikin hanyar polypeptide, saboda steric hani da manyan kungiyoyi suka haifar a cikin kayan ado na gefe, yana da wuya ga phosphorylation na amino acid don raguwa tare da sarkar peptide.A nan gaba, gabatarwar amino acid zai zama mafi wahala, musamman a cikin kayan ado mai dauke da wuraren phosphorylation da yawa, tsararrakin zai zama da wahala sosai, samfurin samfurin ƙarshe yana da wuyar gaske, yana da wuya a rabu, kuma yawan amfanin ƙasa ya ragu sosai.
Saboda haka, lokacin da mahara shafukan a cikin peptide sarkar da aka phosphorylated, wanda zai iya zabar don samar da peptide jerin a kan guduro sa'an nan phosphorylate shi ta gefen sarkar hydroxyl kungiyar Ser, Tyr, ko Thr: ta tsara yafi ya ƙunshi zaɓaɓɓen kau da. ƙungiyar kare sarkar gefe wacce ke buƙatar sanin amino acid bayan haɗin peptide.Ga Tyr, Thr na iya amfani da sarƙoƙin gefen amino acid kai tsaye ba su da kariya ta amsa.
Tushen kariyar sarkar gefe 1% muhallin TFA/DCM na iya cirewa da yawa.Lokacin da aka zaɓi wannan hanyar, ana iya samar da phosphoramidite tetrazolium mai aiki tsaka-tsaki ta amfani da dibenzyl phosphoramidite da tetrazolium, wanda aka haɗa da ƙungiyar hydroxyl, sa'an nan kuma oxidized don samar da ƙungiyar phosphoryl a cikin yanayin peroxyacid don kammala aikin.
Lokacin aikawa: Satumba-08-2023