Isolation and Characterization of Novel Antimicrobial Peptides from the Zebrafish, Danio rerio
EMSL Project ID
2499
Abstract
Antimicrobial peptides have been shown to play a key role in the innate immune defense of a wide variety of organisms, ranging from bacteria to human beings. They have been demonstrated to possess anti-bacterial, anti-fungal, anti-viral, anti-parasitic, and anti-cancer properties. The classical understanding of antimicrobial peptide function involves the interaction of the peptide with a biomembrane, following by disruption and eventual cell death. However, recent evidence indicates that antimicrobial peptides may be able to modulate and affect immune responses in ways more complex than these simple membrane interactions. Indeed, it is our hope that we may begin to examine some of these questions using the zebrafish model system. Antimicrobial peptides have yet to be isolated from the zebrafish, Danio rerio. Typical genetic approaches involving EST database mining have proven difficult due to the lack of conservation at the amino acid level and the relative paucity of fish antimicrobial peptides that have already been identified. As a result, we have chosen to isolate novel antimicrobial peptides via chromatography and mass spectrometry. In preliminary experiments with Dr. Suzana Martinovic, bioactive fractions isolated from strong cation exchange HPLC were further purified using HPLC-MS and HPLC-MS/MS. In one such experiment, over 5000 masses were detected. As a result, we would respectfully request further collaboration with your laboratory. We hope to characterize other bioactive fractions using HPLC-MS and HPLC-MS/MS techniques and to develop a zebrafish database from which we can assign identities to specific data points. We ask permission for time on the 7T FT-ICR MS instrument and help in the development and use of a zebrafish database. The sensitivity of the FT-ICR MS instruments will be necessary for analysis of our samples because of the small sample amounts and low concentrations.
Project Details
Project type
Exploratory Research
Start Date
2002-06-03
End Date
2003-05-19
Status
Closed
Released Data Link
Team
Principal Investigator
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