He kuleana koʻikoʻi ia i nā kaʻina ola, phosphorylating Tyrr, Ser, Thr, aia ma nā peptides.Hoʻohana mau ʻia ka waikawa amino phosphorylated i ka hana ʻana i ka peptide phosphoric acid, i kēia manawa i hoʻohana ʻia i loko o ka phenyl phosphorylated amino acids.Hoʻohana maʻamau ʻia ka HBTU / phosphorylated amino acids e hoʻopili ai i ka phosphorylated amino acids HOBt/DIEA, akā ke koho nei ka phosphoric acid peptide i kēia ala i nā hemahema, ʻoi aku hoʻi i nā peptide peptide lōʻihi i hana ʻia a i ʻole amino acid phosphate, ka hoʻopili ʻana i ka hana haʻahaʻa, haʻahaʻa. ka maemae o ka hua hope.No kēia phosphopeptide, ua noʻonoʻo mākou i kahi ala post-phosphorylation koho.ʻO ke kaʻina hana ʻo ia ka wehe ʻana i nā pūʻulu pale ʻaoʻao-chain amino acid e pono e hōʻailona ʻia ma hope o ka synthesis peptide.No Tyr, hiki iā Thr ke pane pololei me ka hoʻohana ʻana i nā waikawa amino me nā kaulahao ʻaoʻao ʻaoʻao ʻole.-Ser (trt), wehe nui ʻia ma 1%TFA /DCM kaiapuni.
Phosphorylation, me papalua benzene phosphite phosphoramide a me eha naikokene ua hanaia a me ka phosphoramide tetrazolium ikaika intermediates, pili i ka hydroxy, a laila ma lalo o ka waika peroxide oxidation phosphoryl hoʻokumu, piha hopena.
I kēia manawa, ʻelua mau ʻano nui o ka peptide phosphorylated modification method:
(1) ka hoʻokomo pololei ʻana i nā waikawa amino phosphorylated i hoʻopaʻa ʻia i loko o ke kaʻina polypeptide;
(2) ma hope o ka resin i hana ʻia ma ke kaʻina peptide, iā Ser, Tyr, a i ʻole Thr phosphorylation o ke kaulahao ʻaoʻao hydroxyl.
1) e hōʻoiaʻiʻo i ka phosphorylated amino acids e hoʻokomo pololei i ke kaʻina peptide:
ʻO ia hoʻi, ʻo nā amino acids e pono e phosphorylated e phosphorylated a mālama pono ʻia ma mua o ka hoʻohana ʻana (Thr, Ser or Tyr), a laila e hōʻemi ʻia ka phosphorylation i kahi koho o ka index polypeptide e pili ana i ke kaʻina hana SPPS maʻamau. .He maʻalahi kēia ʻano hana, ua lilo ia i kumu nui o nā ʻoihana peptide phosphorylated hoʻololi.
2) ma hope o ka resin i hana ʻia ma ke kaʻina peptide, iā Ser hydroxy phosphorylation, Tyr, a i ʻole Thr mau kaulahao ʻaoʻao:
ʻO ka hoʻonaninani Phosphorylation me ka hoʻohana ʻana i ka phosphorylation o ka hoʻokaʻawale hoʻokahi i ke ʻano polypeptide, ma muli o ke keʻakeʻa steric i hana ʻia e nā hui nui i ka hoʻonaninani ʻaoʻao ʻaoʻao, paʻakikī i ka phosphorylation o nā waikawa amino e hoʻemi me ke kaulahao peptide.I ka wā e hiki mai ana, ʻoi aku ka paʻakikī o ka hoʻokomo ʻana i nā waikawa amino, ʻoi aku ka paʻakikī o ka hoʻonaninani ʻana i nā pūnaewele phosphorylation he nui, e lilo ka hanauna i mea paʻakikī loa, paʻakikī ka hoʻohui ʻana o ka huahana hope, paʻakikī ke hoʻokaʻawale, a haʻahaʻa loa ka hua.
No laila, i ka wā e phosphorylated ai nā wahi he nui i ke kaulahao peptide, hiki i kekahi ke koho e hoʻohua i ke kaʻina peptide ma ka resin a laila phosphorylate iā ia e ka hui hydroxyl ʻaoʻao o Ser, Tyr, a i ʻole Thr: ʻo kāna hana nui e pili ana i ka wehe ʻana o ka ʻO ka hui pale kaulahao ʻaoʻao e pono ai ka ʻike ʻana i nā waikawa amino ma hope o ka synthesis peptide.No Tyr, hiki iā Thr ke hoʻohana pololei i nā kaulahao ʻaoʻao amino ʻaʻole i pale ʻia i ka pane.
ʻO ke kahua palekana kaulahao ʻaoʻao 1% TFA/DCM kaiapuni hiki ke hoʻoneʻe nui ʻia.Ke koho ʻia kēia ʻano hana, hiki ke hana ʻia kahi phosphoramidite tetrazolium active intermediate me ka hoʻohana ʻana i ka dibenzyl phosphoramidite a me tetrazolium, i hoʻopili ʻia i kahi pūʻulu hydroxyl, a laila oxidized e hana i kahi hui phosphoryl i loko o ka peroxyacid environment e hoʻopau ai i ka hopena.
Ka manawa hoʻouna: Sep-08-2023