P450 enzymes identified using LC MS/MS data and database searching with Mascot
Microsomal proteins were resolved using SDS-PAGE, and gel bands 7, 8, and 9 were used for identification and relative quantification of P450 enzymes. The databases used for identification were Swissprot, MSDB, trEMBL, and NCBInr. The best proteotypic peptides were used to confirm the identified proteins and can be used for absolute quantification with AQUA or QconCAT. These peptides were unique for each of the proteins and were selected to ensure complete digestion with trypsin and limited modifications for highly reactive amino acid residues (cysteines and methionines).
P450 Enzyme | ExPasy Accession Code | NCBI Accession Number | Mr | Mascot Score | emPAI | Number of Peptides Identified | Number of Unique Peptides | Sequences of the Best Proteotypic Peptides |
---|---|---|---|---|---|---|---|---|
Da | ||||||||
CYP1A2 | gi22730405 | 58,238 | 40 | 0.06 | 1 | 1 | NSHEFVETASSGNPVDFFPILR | |
CYP2A19 | Q8SQ68 | gi19071834 | 56,857 | 345 | 0.88 | 11 | 8 | DFIDSFLIR, GEQATFDWLFK, GAFIDPTYFLSR, SPQLPQDIDVSPK, EALVDQAEEFSGR, IQEEAGYLIEAFR |
CYP2C33v4 | Q8SQ66 | gi47523878 | 56,634 | 140 | 0.13 | 3 | 2 | EALIDQGDEFLGR, YIALLPSNLPHATVR |
CYP2C49 | Q8SQ65 | gi47523894 | 55,439 | 126 | 0.29 | 4 | 2 | FTLEPVVDLK, EALIDGGEEFSGR, FDYTDPNFLTLLEK, LPPGPTPLSILGNILQLDVK |
CYP2D25 | O46658 | gi47523842 | 56,875 | 381 | 0.66 | 9 | 7 | LLDLVLEGLK, GTTLITNLTSVLK, GNPESSFNDENLR, DLAQPPR |
CYP2E1 | Q8SQ64 | gi47523896 | 57,095 | 114 | 0.21 | 4 | 1 | FIDLIPSNLPHEATR, LPPGPFPLPIVGNIFQLDLK, GTVVIPTLDSVLYDSQEFPEPEK, FIDLIPSNLPHEATR, DIDLSPIAIGFAK |
CYP3A29 | P79401 | gi47523900 | 57,199 | 116 | 0.21 | 3 | 1 | NALSLAEDEEWK, LQEEIEATFPNK, DTINPYTYLPFGTGPR |
CYP3A39 | Q8SQ59 | gi47523898 | 57,137 | 45 | 0.15 | 2 | 1 | LQEEIDATFPSK |
CYP4A21 | Q9GJX5 | gi40781678 | 57,635 | 44 | 0.21 | 5 | 4 | SAFLQNDIIYR, ELSKPITFPDGR, EFQEESELQPLLK |
CYP51A1 | D0G6S0 | gi11066260 | 56,876 | 58 | 0.08 | 1 | 1 | IDDILQTLLDSTYK, YLQDNPASGEK |