The ability to sequence and identify proteins, map their sites of post-translational modification (PTM), and assess their abundances is central to modern biology. Mass spectrometry (MS) is the gold standard technology by which this information is obtained. Serving as the centerpiece, tandem MS (MS/MS) is a principal component. Electron transfer dissociation (ETD), a relatively new MS/MS dissociation method, has generated significant excitement for its compatibility with previously intractable peptide/protein classes. Five years ago m/z range, mass accuracy, and mass resolution considerably restricted the application of ETD. Our initial RO1 proposal successfully eliminated this limitation by coupling ETD to the orbitrap mass analyzer. The resulting system routinely analyzes peptides and proteins, with and without labile PTMs, with a high-fidelity readout (orbitrap). As a result, it realized many of our anticipated outcomes and created numerous unforeseen opportunities. Just in the PI's laboratory, the latter set includes data-dependent selection of dissociation method (i.e., Decision Tree), discovery of the unique chemical compositions of z-type ions, internal spectral calibration using ETD reagents, activated-ion ETD, and several biological applications. By 2008, the commercial implementation of our technology began to reach researchers across the globe-nearly 300 to date-enabling access to numerous previously intractable problems such as mapping Arg methylation sites, increasing coverage of low molecular weight proteins, providing unambiguous PTM site assignment, and screening glycopeptide libraries, among many others. We detail two new aims that build upon the high impact results of our initial funding period.
Aim 1, how do we broaden the utility of ETD for biomedical research? Aim 2, what is the role of gas- phase purification in quantitative proteomics? We continue with a balance of instrumentation, method, informatic, and applied projects constructed upon the widely used ETD-orbitrap platform we described 3.5 years ago.
Cutting edge MS based technology, Electron transfer dissociation (ETD), continues to be developed. This new MS/MS dissociation method enables previously intractable peptide/protein classes to be sequenced and identified, have their sites of post-translational modification (PTM) mapped, and assess their abundances. This is central to modern biology and has relevance for research ranging from human disease to evolution.
| Lera, Robert F; Potts, Gregory K; Suzuki, Aussie et al. (2016) Decoding Polo-like kinase 1 signaling along the kinetochore-centromere axis. Nat Chem Biol 12:411-8 |
| Wilkerson, Emily M; Johansson, Mats W; Hebert, Alexander S et al. (2016) The Peripheral Blood Eosinophil Proteome. J Proteome Res 15:1524-33 |
| Baughman, Joshua M; Rose, Christopher M; Kolumam, Ganesh et al. (2016) NeuCode Proteomics Reveals Bap1 Regulation of Metabolism. Cell Rep 16:583-595 |
| Horton, Julie L; Martin, Ola J; Lai, Ling et al. (2016) Mitochondrial protein hyperacetylation in the failing heart. JCI Insight 2: |
| Riley, Nicholas M; Coon, Joshua J (2016) Phosphoproteomics in the Age of Rapid and Deep Proteome Profiling. Anal Chem 88:74-94 |
| Riley, Nicholas M; Mullen, Christopher; Weisbrod, Chad R et al. (2016) Enhanced Dissociation of Intact Proteins with High Capacity Electron Transfer Dissociation. J Am Soc Mass Spectrom 27:520-31 |
| Potts, Gregory K; Voigt, Emily A; Bailey, Derek J et al. (2016) Neucode Labels for Multiplexed, Absolute Protein Quantification. Anal Chem 88:3295-303 |
| Ledvina, Aaron R; Coon, Joshua J; Ture?ek, František (2015) Competitive Hydrogen Atom Migrations Accompanying Cascade Dissociations of Peptide Cation-Radicals of the z (+•) Type. Int J Mass Spectrom 377:44-53 |
| Rhoads, Timothy W; Prasad, Aman; Kwiecien, Nicholas W et al. (2015) NeuCode Labeling in Nematodes: Proteomic and Phosphoproteomic Impact of Ascaroside Treatment in Caenorhabditis elegans. Mol Cell Proteomics 14:2922-35 |
| Richards, Alicia L; Hebert, Alexander S; Ulbrich, Arne et al. (2015) One-hour proteome analysis in yeast. Nat Protoc 10:701-14 |
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