Subcellular fractionation of human liver reveals limits in global proteomic quantification from isolated fractions.

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  • Additional Information
    • Source:
      Publisher: Elsevier Country of Publication: United States NLM ID: 0370535 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1096-0309 (Electronic) Linking ISSN: 00032697 NLM ISO Abbreviation: Anal Biochem Subsets: MEDLINE
    • Publication Information:
      Publication: <2000- > : San Diego, CA : Elsevier
      Original Publication: Orlando Fl : Academic Press
    • Subject Terms:
      Cell Fractionation/*methods ; Liver/*chemistry ; Proteome/*chemistry ; Proteomics/*methods; Female ; Humans ; Male
    • Abstract:
      The liver plays an important role in metabolism and elimination of xenobiotics, including drugs. Determination of concentrations of proteins involved in uptake, distribution, metabolism, and excretion of xenobiotics is required to understand and predict elimination mechanisms in this tissue. In this work, we have fractionated homogenates of snap-frozen human liver by differential centrifugation and performed quantitative mass spectrometry-based proteomic analysis of each fraction. Concentrations of proteins were calculated by the "total protein approach". A total of 4586 proteins were identified by at least five peptides and were quantified in all fractions. We found that the xenobiotics transporters of the canalicular and basolateral membranes were differentially enriched in the subcellular fractions and that phase I and II metabolizing enzymes, the cytochrome P450s and the UDP-glucuronyl transferases, have complex subcellular distributions. These findings show that there is no simple way to scale the data from measurements in arbitrarily selected membrane fractions using a single scaling factor for all the proteins of interest. This study also provides the first absolute quantitative subcellular catalog of human liver proteins obtained from frozen tissue specimens. Our data provide quantitative insights into the subcellular distribution of proteins and can be used as a guide for development of fractionation procedures.
      (Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.)
    • Contributed Indexing:
      Keywords: Absolute quantitative proteomics; Drug metabolism; Drug transport; Human liver; Subcellular fractionation; Total protein approach
    • Accession Number:
      0 (Proteome)
    • Publication Date:
      Date Created: 20160618 Date Completed: 20170428 Latest Revision: 20180207
    • Publication Date:
      20240829
    • Accession Number:
      10.1016/j.ab.2016.06.006
    • Accession Number:
      27311553