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Diagnosis of Ecosystem Adaptive Responses by Analysis of Hundreds of Peptide Biomarkers in the North Atlantic Ocean using Targeted Metaproteomics

Reputable Mentor II
Reputable Mentor II
Mak Saito1, Matt McIlvin1, Dawn Moran1, Luis Valentin1, Romain Huguet2, Graeme McAlister2, Shannon Eliuk2, Rod Johnson3
ASMS 2017 Poster
Motivation ● Ocean microbes are Earth’s life support system ● Developing Targeted metaproteomics to detect and diagnose long-term ecosystem and biogeochemical changes ● Absolute concentrations are necessary because biological matrix is not constant ● Proof-of-concept at monthly sampling of the Bermuda Atlantic Time Series Station Methodologies and Results ● Ship-based submersible pump sample collection of ~300L/sample ● Magnetic bead single pot extraction (Hughes et al., 2014) ● 2D active modulation discovery proteomics on Thermo Fusion ● Coupling to metagenomic databases for PSM ● Standard production with 6+ carousels of QConCat 15N labeled peptides, externally calibrated ● Parallel Reaction Monitoring of 157 tryptic peptides (314 including heavies) using QuanDirect on Thermo Scientific Orbitrap Fusion MS ● PRM Peak analysis in Skyline ● Taxon attribution of tryptic peptides in METATRYP (Saito et al., Proteomics 2015) ● 77% of the targeted peptides (122 of 157) gave positive values in 3 or more of the seven samples in a vertical profile

1 Marine Chemistry and Geochemistry Department, Woods Hole Oceanographic Institution 2 Thermo Fisher Scientific San Jose 3 Bermuda Institute of Ocean Sciences
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Last update:
‎10-15-2021 11:31 AM
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