Proteome Profile of Cytosolic Component of Zebrafish Liver Generated by LC-ESI MS/MS Combined With Trypsin Digestion and Microwave-Assisted Acid Hydrolysis

J Proteome Res. 2007 Jan;6(1):263-72. doi: 10.1021/pr060367o.

Abstract

The zebrafish genome has recently been sequenced and annotated allowing for high-throughput proteomic analysis. Here, we report for the first time a proteomic subset of zebrafish liver, an important organ for metabolizing toxins. Using a newly developed analytical procedure, we have identified 1204 proteins from the cytosolic component of a zebrafish liver tissue sample. Our methods involve cell-compartment fractionation of liver tissue samples, four levels of protein digestion, and off-line two-dimensional liquid chromatography (2-D LC) separations of resultant peptides. Proteins are identified using an electrospray ionization quadrupole time-of-flight tandem mass spectrometer (ESI-QTOF MS/MS), which provides high-resolution and high-accuracy mass measurement of peptide ions and their fragment ions. We demonstrate that greater proteome coverage can be achieved by combining the results obtained from four methods of protein digestion: three tryptic digests (one in buffer, one in methanol, and another in SDS), and a microwave-assisted acid hydrolysate of the protein extracts. Identified proteins--which included several groups of established protein biomarkers--were functionally classified. We discuss the functions and implications of these biomarkers within the context of zebrafish toxicology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatography, Ion Exchange
  • Cytosol / metabolism*
  • Hydrolysis
  • Liver / metabolism*
  • Microwaves*
  • Peptide Mapping / methods
  • Proteome
  • Proteomics / methods*
  • Spectrometry, Mass, Electrospray Ionization
  • Subcellular Fractions
  • Time Factors
  • Trypsin / pharmacology*
  • Zebrafish

Substances

  • Proteome
  • Trypsin