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

[Orbitrap_SciLib]

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