Expanding the zebrafish embryo proteome using multiple fractionation approaches and tandem mass spectrometry
- Authors
- Lößner, C., Wee, S., Ler, S.G., Li, R.H., Carney, T., Blackstock, W., and Gunaratne, J.
- ID
- ZDB-PUB-120604-6
- Date
- 2012
- Source
- Proteomics 12(11): 1879-1882 (Journal)
- Registered Authors
- Carney, Tom
- Keywords
- deyolking, fractionation, gene ontology, whole proteome, zebrafish embryo
- MeSH Terms
-
- Animals
- Cell Fractionation/methods
- Extracellular Space/chemistry
- Nucleocytoplasmic Transport Proteins/chemistry
- Proteome/analysis*
- Proteomics
- Ribosomal Proteins/chemistry
- Tandem Mass Spectrometry
- Zebrafish/embryology*
- Zebrafish Proteins/analysis*
- PubMed
- 22653788 Full text @ Proteomics
The proteome of zebrafish, Danio rerio, embryos has not been studied in great detail mainly due to the presence of high abundance yolk proteins in embryos. Here we report the highest number of the zebrafish embryo proteins identified so far to our knowledge, through a combination of a protein level fractionation approach (1D SDS-PAGE) and two different peptide level fractionation approaches (isoelectric focusing (IEF) and strong anion exchange (SAX)) of deyolked zebrafish embryos followed by LC-MS/MS. We detected 5,267 proteins in total of which 3,464 proteins were identified with at least two peptides (less than 1% peptide false discovery rate). The analysis of proteome coverage from each method showed that 56% of detected proteins were common to all approaches and 95% of the detected proteome was obtained from 1D SDS PAGE approach alone. Bioinformatics analysis of the detected proteome demonstrated that nucleocytoplasmic transport (biological process) and ribosomal proteins (cellular component) were the most over-represented proteins, whereas cell-cell signaling (biological process) and extracellular space proteins (cellular component) were the most under-represented proteins in the identified proteome.