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A gel-free MS-based quantitative proteomic approach accurately measures cytochrome P450 protein concentrations in human liver microsomes.

Publication ,  Journal Article
Wang, MZ; Wu, JQ; Dennison, JB; Bridges, AS; Hall, SD; Kornbluth, S; Tidwell, RR; Smith, PC; Voyksner, RD; Paine, MF; Hall, JE
Published in: Proteomics
October 2008

The human cytochrome P450 (P450) superfamily consists of membrane-bound proteins that metabolize a myriad of xenobiotics and endogenous compounds. Quantification of P450 expression in various tissues under normal and induced conditions has an important role in drug safety and efficacy. Conventional immunoquantification methods have poor dynamic range, low throughput, and a limited number of specific antibodies. Recent advances in MS-based quantitative proteomics enable absolute protein quantification in a complex biological mixture. We have developed a gel-free MS-based protein quantification strategy to quantify CYP3A enzymes in human liver microsomes (HLM). Recombinant protein-derived proteotypic peptides and synthetic stable isotope-labeled proteotypic peptides were used as calibration standards and internal standards, respectively. The lower limit of quantification was approximately 20 fmol P450. In two separate panels of HLM examined (n = 11 and n = 22), CYP3A, CYP3A4 and CYP3A5 concentrations were determined reproducibly (CV

Duke Scholars

Published In

Proteomics

DOI

EISSN

1615-9861

ISSN

1615-9853

Publication Date

October 2008

Volume

8

Issue

20

Start / End Page

4186 / 4196

Related Subject Headings

  • Uncertainty
  • Sequence Homology, Amino Acid
  • Proteomics
  • Middle Aged
  • Microsomes, Liver
  • Mass Spectrometry
  • Male
  • Humans
  • Female
  • Cytochrome P-450 Enzyme System
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Wang, M. Z., Wu, J. Q., Dennison, J. B., Bridges, A. S., Hall, S. D., Kornbluth, S., … Hall, J. E. (2008). A gel-free MS-based quantitative proteomic approach accurately measures cytochrome P450 protein concentrations in human liver microsomes. Proteomics, 8(20), 4186–4196. https://doi.org/10.1002/pmic.200800144
Wang, Michael Zhuo, Judy Qiju Wu, Jennifer B. Dennison, Arlene S. Bridges, Stephen D. Hall, Sally Kornbluth, Richard R. Tidwell, et al. “A gel-free MS-based quantitative proteomic approach accurately measures cytochrome P450 protein concentrations in human liver microsomes.Proteomics 8, no. 20 (October 2008): 4186–96. https://doi.org/10.1002/pmic.200800144.
Wang MZ, Wu JQ, Dennison JB, Bridges AS, Hall SD, Kornbluth S, et al. A gel-free MS-based quantitative proteomic approach accurately measures cytochrome P450 protein concentrations in human liver microsomes. Proteomics. 2008 Oct;8(20):4186–96.
Wang, Michael Zhuo, et al. “A gel-free MS-based quantitative proteomic approach accurately measures cytochrome P450 protein concentrations in human liver microsomes.Proteomics, vol. 8, no. 20, Oct. 2008, pp. 4186–96. Epmc, doi:10.1002/pmic.200800144.
Wang MZ, Wu JQ, Dennison JB, Bridges AS, Hall SD, Kornbluth S, Tidwell RR, Smith PC, Voyksner RD, Paine MF, Hall JE. A gel-free MS-based quantitative proteomic approach accurately measures cytochrome P450 protein concentrations in human liver microsomes. Proteomics. 2008 Oct;8(20):4186–4196.
Journal cover image

Published In

Proteomics

DOI

EISSN

1615-9861

ISSN

1615-9853

Publication Date

October 2008

Volume

8

Issue

20

Start / End Page

4186 / 4196

Related Subject Headings

  • Uncertainty
  • Sequence Homology, Amino Acid
  • Proteomics
  • Middle Aged
  • Microsomes, Liver
  • Mass Spectrometry
  • Male
  • Humans
  • Female
  • Cytochrome P-450 Enzyme System