PUBLICATION
Proteome Profile of Cytosolic Component of Zebrafish Liver Generated by LC-ESI MS/MS Combined with Trypsin Digestion and Microwave-Assisted Acid Hydrolysis
- Authors
- Wang, N., Mackenzie, L., Souza, A.G., Zhong, H., Goss, G., and Li, L.
- ID
- ZDB-PUB-070122-17
- Date
- 2007
- Source
- Journal of Proteome Research 6(1): 263-272 (Journal)
- Registered Authors
- Goss, Greg
- Keywords
- zebrafish, toxicology, biomarker, liver proteome, LC-ESI MS/MS, microwave-assisted acid hydrolysis
- 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
- PubMed
- 17203970 Full text @ J. Proteome Res.
Citation
Wang, N., Mackenzie, L., Souza, A.G., Zhong, H., Goss, G., and Li, L. (2007) Proteome Profile of Cytosolic Component of Zebrafish Liver Generated by LC-ESI MS/MS Combined with Trypsin Digestion and Microwave-Assisted Acid Hydrolysis. Journal of Proteome Research. 6(1):263-272.
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&sbd;which included several groups of established protein biomarkers&sbd;were functionally classified. We discuss the functions and implications of these biomarkers within the context of zebrafish toxicology.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping