Zogulitsa

Kusanthula kwa zochitika za proteome-wide phosphorylation akadali vuto lalikulu lowunikira chifukwa chazovuta kwambiri zama protein phosphorylation network.Mu ntchitoyi, tikuwunika kugwirizana kwa Lys-N, Lys-C, ndi trypsin potengera kuthekera kwawo kothandizira kuwunika kwa phosphoproteome padziko lonse lapansi.Mtundu woyengedwa wa kusinthana kwamphamvu kwa pH ya pH idagwiritsidwa ntchito kulekanitsa bwino ma peptides a N-terminal acetylated, phosphorylated, ndi nonmodified.Chiwerengero cha 5036 phosphopeptides chosagwiritsidwa ntchito chikhoza kudziwika ndi chiwerengero chabodza cha <1% kuchokera ku 1 mg ya mapuloteni pogwiritsa ntchito ma enzyme atatu.Deta yathu idawulula kuti kuphatikizika pakati pa ma data a phosphopeptide opangidwa ndi ma protease osiyanasiyana kunali kocheperako, pomwe kuphatikizika pakati pa seti ziwiri zofananira zama data za tryptic zidapezeka kuti ndizosachepera nthawi za 4.Mwanjira iyi, kugwiritsidwa ntchito kofanana kwa Lys-N ndi trypsin kunapangitsa kuti chiwonjezeko cha 72% chiwonjezeko cha ma phosphopeptides odziwika poyerekeza d ndi trypsin okha, pomwe kuyesa koyerekeza kwa trypsin kunangowonjezera 25%.Chifukwa chake, poyang'ana kwambiri za trypsin ndi Lys-N, tidazindikira ma phosphopeptides 4671 osafunikira.Kusanthula kwina kwa malo omwe apezeka kunawonetsa kuti ma data a Lys-N ndi trypsin adalemeretsedwa mumitundu yosiyana kwambiri ya phosphorylation, kutsimikiziranso kuti njira zama multiprotease ndizofunika kwambiri pakuwunika kwa phosphoproteome.