He whai waahi nui ki roto i nga mahi ora, ko te phosphorylating Tyrr, Ser, Thr, kei runga i nga peptides.Ko te waikawa amino phosphorylated e whakamahia nuitia ana ki te hanga peptide waikawa phosphoric, e whakamahia ana i tenei wa i roto i nga waikawa amino phenyl phosphorylated kotahi.HBTU / phosphorylated amino acids e whakamahia nuitia ana ki te hono i nga waikawa amino phosphorylated HOBt / DIEA tikanga, engari ko te phosphoric acid peptide kei te kowhiri i tenei huarahi koha, ina koa i roto i nga peptide peptide roa ake, te waikawa amino phosphate ranei, he iti te pai o te hono, he iti. te ma o te hua whakamutunga.Mo tenei phosphopeptide, i whakaarohia e matou he huarahi whiriwhiri i muri i te phosphorylation.Ko te tukanga whakatipuranga ko te tango i nga roopu whakamarumaru taha-taha waikawa amino e tika ana kia tapaina i muri i te whakahiato peptide.Mo Tyr, ka taea e Thr te urupare tika ma te whakamahi i nga waikawa amino me nga mekameka taha kore.-Ser (trt), kua tangohia ine i te 1%TFA /DCM taiao.
Phosphorylation, ki te rua benzene phosphite phosphoramide me wha hauota e hanga me te phosphoramide tetrazolium kaha takawaenga, hono ki te hydroxy, ka i raro i te waikawa peroxide phosphoryl hanganga, oti tauhohenga.
I tenei wa, ko te tikanga whakarereke phosphorylated peptide e rua nga momo matua:
(1) te whakauru tika i nga waikawa amino phosphorylated ki roto i te raupapa polypeptide;
(2) i muri i te kapia i hangaia i runga i te raupapa peptide, ki Ser, Tyr, Thr phosphorylation ranei o te taha mekameka hydroxyl.
1) ka whakatauhia nga waikawa amino phosphorylated ki te whakauru tika i te raupapa peptide:
Arā, ko nga waikawa amino e tika ana kia phosphorylated ka phosphorylated me te tiaki tika i mua i te whakamahi (Thr,Ser or Tyr), katahi ka whakawhäitihia te phosphorylation ki te waahi whiriwhiri o te taupū polypeptide i runga i te tikanga whakangao SPPS noa. .He ngawari tenei tikanga ki te whakahaere, kua waiho hei tikanga matua mo nga umanga peptide phosphorylated whakarereke.
2) i muri i te kapia i hangaia i runga i te raupapa peptide, ki te Ser hydroxy phosphorylation, Tyr, Thr ranei nga mekameka taha:
Te whakapaipai o te Phosphorylation ma te whakamahi i te phosphorylation o te whakahekenga kotahi ki roto i te tikanga polypeptide, na te aukati steric i hangaia e nga roopu nui i roto i te whakapaipai mekameka taha, he uaua mo te phosphorylation o nga waikawa amino ki te whakaheke me te mekameka peptide.I te wa kei te heke mai, ka uaua ake te whakauru o nga waikawa amino, ina koa i roto i te whakapaipai kei roto i nga waahi phosphorylation maha, ka tino uaua te whakatipuranga, he uaua te hanganga o te hua whakamutunga, he uaua ki te wehe, he iti rawa te hua.
No reira, ka phosphorylated nga waahi maha i roto i te mekameka peptide, ka taea e tetahi te kowhiri ki te whakaputa i te raupapa peptide i runga i te kapia katahi ka whakapourihia e te roopu hydroxyl mekameka taha o Ser, Tyr, Thr ranei: ko tana whakatipuranga te nuinga o te tango i te tangohanga o te Ko te roopu tiaki mekameka taha e hiahia ana kia mohiohia nga waikawa amino i muri i te whakahiato peptide.Mo Tyr, ka taea e Thr te whakamahi tika i nga mekameka taha waikawa amino kaore e tiakina hei whakautu.
Te turanga tiaki mekameka taha 1% TFA/DCM taiao ka taea te tango ine.Ina whiriwhiria tenei tikanga, ka taea te hanga i tetahi takawaenga kaha phosphoramidite tetrazolium ma te whakamahi i te dibenzyl phosphoramidite me te tetrazolium, honoa ki te roopu hydroxyl, katahi ka oxidized ki te hanga i te roopu phosphoryl i roto i te taiao peroxyacid hei whakaoti i te tauhohenga.
Wā tuku: Sep-08-2023