Comprehensive identification of cytochrome p450 isoforms from solubility-based fractionated rat liver microsomes.
Cytochrome P450 isoforms from male rat liver microsomes were comprehensively identified using nano liquid chromatography tandem mass spectrometry (nanoLC-MS/MS). The enrichment of P450, an endomembrane-anchored heme protein, was achieved by solubility-based protein fractionation, and greatly improved the total number of identified P450 isoforms. LC-MS/MS analysis of fractions resulted in the identification of total 36 P450 isoforms. The combination of proteomic analysis and the solubility-based fractionation would provide powerful tool for the expression analysis of the superfamily proteins having great similarities between the amino acids sequences.
Chemicals Assessment Center, Chemicals Evaluation and Research Institute, 1600, Shimo-Takano, Sugito-machi, Kitakatsushika-gun, Saitama 345-0043, Japan. yamanaka-hidenori@ceri.jp
Hidenori
H
Yamanaka
Masahiro
M
Takeyoshi
Yasushi
Y
Minobe
Yoshikuni
Y
Yakabe
Mineo
M
Takatsuki
Hisaya
H
Sato
D000818:Animals; D005591:Chemical Fractionation; D002853:Chromatography, Liquid / Q000379:methods*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D036103:Nanotechnology / Q000379:methods; D051381:Rats; D017207:Rats, Sprague-Dawley; D012995:Solubility; D053719:Tandem Mass Spectrometry / Q000379:methods
D007527:Isoenzymes; D003577:Cytochrome P-450 Enzyme System