Nā huahana

ʻO ka nānā ʻana i nā hanana phosphorylation proteome-ākea he paʻakikī koʻikoʻi nui no ka paʻakikī nui o nā pūnaewele phosphorylation protein.Ma kēia hana, loiloi mākou i ka hoʻohui ʻana o Lys-N, Lys-C, a me trypsin e pili ana i ko lākou hiki ke hāʻawi i ka loiloi honua o ka phosphoproteome.Ua hoʻohana ʻia kahi mana hoʻomaʻemaʻe o ka hoʻololi cation ikaika haʻahaʻa pH no ka hoʻokaʻawale maikaʻi ʻana i nā peptides N-terminally acetylated, phosphorylated, a me nā peptides nonmodified.Hiki ke ʻike ʻia ka huina o 5036 nonredundant phosphopeptides me ka helu ʻike wahaheʻe o <1% mai ka 1 mg o ka protein e hoʻohana ana i ka hui ʻana o nā enzymes ʻekolu.Ua hōʻike ʻia kā mākou ʻikepili ʻo ka overlap ma waena o nā pūʻulu data phosphopeptide i hana ʻia me nā protease ʻokoʻa he marginal, ʻoiai ʻo ka overlap ma waena o ʻelua mau pūʻulu ʻike tryptic i hana like ʻia ua ʻike ʻia ma kahi o 4 mau manawa kiʻekiʻe.Ma kēia ala, hiki i ka hoʻohana like ʻana o Lys-N a me trypsin ke hoʻonui i ka 72% o ka nui o nā phosphopeptides i ʻike ʻia e like me ka hoʻohālikelike ʻana i ka trypsin wale nō, ʻoiai kahi hoʻokolohua hoʻohālikelike trypsin i alakaʻi i kahi piʻi 25%.No laila, i ka nānā wale ʻana i ka ʻike trypsin a me Lys-N, ua ʻike mākou i ka 4671 nonredundant phosphopeptides.Ua hōʻike hou ʻia ka nānā ʻana o nā pūnaewele i ʻike ʻia ua hoʻonui ʻia nā ʻikepili ʻo Lys-N a me trypsin i nā kumu phosphorylation ʻē aʻe, e hōʻike hou ana he waiwai nui nā ala multiprotease i nā loiloi phosphoproteome.