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Amine-reactive Neutron-encoded Labels for Highly Plexed Proteomic Quantitation
Journal article   Open access   Peer reviewed

Amine-reactive Neutron-encoded Labels for Highly Plexed Proteomic Quantitation

Alexander S Hebert, Anna E Merrill, Jonathan A Stefely, Derek J Bailey, Craig D Wenger, Michael S Westphall, David J Pagliarini and Joshua J Coon
Molecular & cellular proteomics, Vol.12(11), pp.3360-3369
11/2013
DOI: 10.1074/mcp.M113.032011
PMCID: PMC3820946
PMID: 23882030
url
https://doi.org/10.1074/mcp.M113.032011View
Published (Version of record) Open Access

Abstract

We describe a novel amine-reactive chemical label that exploits differential neutron-binding energy between 13 C and 15 N isotopes. These neutron-encoded (NeuCode) chemical labels enable up to 12-plex MS1-based protein quantification. Each structurally identical, but isotopically unique, tag is encoded with a 12.6-mDa mass difference—relative to its nearest neighbor—so that peptides bearing these NeuCode signatures do not increase spectral complexity and are detected only upon analysis with very high mass-resolving powers. We demonstrate that the method provides quantitative performance that is comparable to both metabolic labeling and isobaric tagging while combining the benefits of both strategies. Finally, we employ the tags to characterize the proteome of Saccharomyces cerevisiae during the diauxic shift, a metabolic transition from fermentation to aerobic respiration.
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