Producing a one-dimensional proteomic map for human liver cytochromes p450.
In this chapter we explore the inducible cytochrome P450 (CYP) forms as an example of membrane proteins analysis that relies on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) fractionation with subsequent mass spectrometric (MS) identification. The approach involves cutting an SDS-PAGE gel lane into thin slices and identifying proteins in each slice by MS with the aim of obtaining detailed information on proteins of interest. A one-dimensional proteomic map showing the distribution of selected CYP isoforms across 40 slices was constructed using mass spectra obtained from each slice. Our protocol proved to be efficient enough to obtain a comprehensive profile of drug-metabolizing enzymes in the human liver. In addition to human tissues, the approach described should be applicable to the characterization of membrane proteins in other eukaryotic species.
Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, Moscow, Russia. cyp450@mail.ru
Natalia A
NA
Petushkova
Andrey V
AV
Lisitsa
D000595:Amino Acid Sequence; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008099:Liver / Q000201:enzymology*; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008862:Microsomes, Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry; D010449:Peptide Mapping; D059748:Proteolysis; D020543:Proteome / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000737:chemistry
D008565:Membrane Proteins; D010446:Peptide Fragments; D020543:Proteome; D003577:Cytochrome P-450 Enzyme System; D014357:Trypsin