Iimveliso

Iprotein phosphorylation ilawula imisebenzi eyahlukeneyo yamaselula kwaye idlala indima ebalulekileyo ekuphuhliseni kwangaphambili kwezityalo.Ukuncedisana nokwandisa kuphando lwangaphambili lweprotein phosphorylation kwizithole ze-Arabidopsis siye sasebenzisa enye indlela edibanisa ukutsalwa kweprotheyini phantsi kweemeko ezingezizo ze-denaturing kunye ne-immobilized metal-ion affinity chromatography (IMAC) etyebisayo ye-phosphoproteins engaguqukiyo kwi-Rubisco-depleted extracts, ilandelwa kukuchongwa. usebenzisa i-electrophoresis yejeli emacala amabini (2-DE) kunye nolwelo lwechromatography-tandem mass spectrometry (LC-MS/MS).I-gel ye-trypsin yokugaya kunye nohlalutyo lweendawo ezikhethiweyo ze-gel ezichongiweyo ze-144 ze-phosphorylated peptides kunye neentsalela, apho kuphela i-phosphopeptides eyi-18 kunye ne-8 phosphosites zifunyenwe kwi-PhosPhat 4.0 kunye ne-P3DB Arabidopsis thaliana phosphorylation yogcino lweenkcukacha.Ngaphezu kwesiqingatha se-82 echongiweyo i-phosphoproteins ibandakanyeka kwi-carbohydrate metabolism, i-photosynthesis / ukuphefumula okanye i-oxidative yokuphendula i-mechani sms.Ukutyetyiswa kwe-phosphoproteins engaguqukiyo ngaphambi kwe-2-DE kunye ne-LC-MS/MS ibonakala iphucula ukubonwa kwe-phosphorylated threonine kunye neentsalela ze-tyrosine xa kuthelekiswa neendlela ezisebenzisa ukutyetyiswa kwenqanaba le-peptide, ebonisa ukuba ezi ndlela zimbini ziyahambelana ngokwemigaqo ye-phosphorylation site coverage.Ukuthelekisa iziphumo zezithole ezincinci kunye nezo zifunyenwe ngaphambili kumagqabi e-Arabidopsis esele zikhulile zichonge iiprotheyini ezintlanu ezahlukeneyo zephosphorylated kwezi zicubu, ebonisa amandla obu buchule bokuphanda amandla eprotein phosphorylation ngexesha lophuhliso lwezityalo.