Kayayyaki

Binciken abubuwan da suka faru na phosphorylation mai faɗin proteome har yanzu babban ƙalubale ne na nazari saboda ɗimbin sarƙaƙƙiya na hanyoyin sadarwar furotin phosphorylation.A cikin wannan aikin, muna kimanta haɓakar Lys-N, Lys-C, da trypsin dangane da ikonsu na ba da gudummawa ga nazarin duniya na phosphoproteome.An yi amfani da ingantaccen sigar ƙaramin pH mai ƙarfi mai ƙarfi don raba N-terminally acetylated, phosphorylated, da peptides waɗanda ba a gyara su yadda ya kamata.Ana iya gano jimlar 5036 phosphopeptides marasa ƙarfi tare da ƙimar gano ƙarya na <1% daga 1 MG na furotin ta amfani da haɗin enzymes uku.Bayanan da muka samu sun nuna cewa hatsaniya tsakanin na'urorin bayanan phosphopeptide da aka samar tare da proteases daban-daban ya kasance a gefe, yayin da aka gano tsakanin nau'ikan bayanan tryptic guda biyu da aka samar da su aƙalla sau 4.Ta wannan hanyar, daidaitaccen amfani da Lys-N da trypsin ya ba da damar haɓaka 72% a cikin adadin phosphopeptides da aka gano idan aka kwatanta d da trypsin kaɗai, yayin da gwajin gwajin trypsin ya haifar da haɓaka 25% kawai.Don haka, lokacin da muka mai da hankali kawai kan trypsin da bayanan Lys-N, mun gano 4671 phosphopeptides marasa ƙarfi.Ƙarin bincike na wuraren da aka gano ya nuna cewa bayanan Lys-N da trypsin sun wadatar da su a cikin nau'o'in phosphorylation daban-daban, suna kara tabbatar da cewa hanyoyin da yawa suna da mahimmanci a nazarin phosphoproteome.