ArticleMolecular & cellular proteomics : MCP2011
Online nanoflow multidimensional fractionation for high efficiency phosphopeptide analysis.
Article in Molecular & cellular proteomics : MCP, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 44 papers.
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Who cites it
44 citing papers in PubMed, 102 citations in OpenAlex.
- High Dynamic Range Peptide Mass Spectrometry Using Segmented Precursor Ion Accumulation.Analytical chemistry · 2025Article
- Leveraging HILIC/ERLIC separations for online nanoscale LC-MS/MS analysis of phosphopeptide isoforms from RNA polymerase II C-terminal domain.Journal of chromatography. B, Analytical technologies in the biomedical and life sciences · 2025Article
- Quantitative Cell Proteomic Atlas: Pathway-Scale Targeted Mass Spectrometry for High-Resolution Functional Profiling of Cell Signaling.Journal of proteome research · 2022Article
- Identification and validation of selective deubiquitinase inhibitors.Cell chemical biology · 2021Article
- Phosphorylation of major Porphyromonas gingivalis virulence factors is crucial for their processing and secretion.Molecular oral microbiology · 2021Article
- Online Nanoflow Two-Dimension Comprehensive Active Modulation Reversed Phase-Reversed Phase Liquid Chromatography High-Resolution Mass Spectrometry for Metaproteomics of Environmental and Microbiome Samples.Journal of proteome research · 2021Article
- Deep phosphoproteome analysis of Schistosoma mansoni leads development of a kinomic array that highlights sex-biased differences in adult worm protein phosphorylation.PLoS neglected tropical diseases · 2020Article
- Review of Three-Dimensional Liquid Chromatography Platforms for Bottom-Up Proteomics.International journal of molecular sciences · 2020Review
- Leveraging Compound Promiscuity to Identify Targetable Cysteines within the Kinome.Cell chemical biology · 2019Article
- In vitro analysis of RNA polymerase II elongation complex dynamics.Genes & development · 2019Article
- In vitro assembly and proteomic analysis of RNA polymerase II complexes.Methods (San Diego, Calif.) · 2019Article
- A Chemoproteomic Strategy for Direct and Proteome-Wide Covalent Inhibitor Target-Site Identification.Journal of the American Chemical Society · 2019Article
- ProteomeGenerator: A Framework for Comprehensive Proteomics Based on de Novo Transcriptome Assembly and High-Accuracy Peptide Mass Spectral Matching.Journal of proteome research · 2018Article
- Synthesizing Signaling Pathways from Temporal Phosphoproteomic Data.Cell reports · 2018Article
- MEF2C Phosphorylation Is Required for Chemotherapy Resistance in Acute Myeloid Leukemia.Cancer discovery · 2018Article
- Advances in microscale separations towards nanoproteomics applications.Journal of chromatography. A · 2017Review
- High Sensitivity Quantitative Proteomics Using Automated Multidimensional Nano-flow Chromatography and Accumulated Ion Monitoring on Quadrupole-Orbitrap-Linear Ion Trap Mass Spectrometer.Molecular & cellular proteomics : MCP · 2017Article
- PRMT1-Mediated Translation Regulation Is a Crucial Vulnerability of Cancer.Cancer research · 2017Article
- Interrogating the hidden phosphoproteome.Proteomics · 2017Article
- Phosphoproteomics of colon cancer metastasis: comparative mass spectrometric analysis of the isogenic primary and metastatic cell lines SW480 and SW620.Analytical and bioanalytical chemistry · 2017Article
Corrections and comments
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Authors and funding
8 authors at 3 institutions in 1 country.
Funding
Abstract
Despite intense, continued interest in global analyses of signaling cascades through mass spectrometry-based studies, the large-scale, systematic production of phosphoproteomics data has been hampered in-part by inefficient fractionation strategies subsequent to phosphopeptide enrichment. Here we explore two novel multidimensional fractionation strategies for analysis of phosphopeptides. In the first technique we utilize aliphatic ion pairing agents to improve retention of phosphopeptides at high pH in the first dimension of a two-dimensional RP-RP. The second approach is based on the addition of strong anion exchange as the second dimension in a three-dimensional reversed phase (RP)-strong anion exchange (SAX)-RP configuration. Both techniques provide for automated, online data acquisition, with the 3-D platform providing the highest performance both in terms of separation peak capacity and the number of unique phosphopeptide sequences identified per μg of cell lysate consumed. Our integrated RP-SAX-RP platform provides several analytical figures of merit, including: (1) orthogonal separation mechanisms in each dimension; (2) high separation peak capacity (3) efficient retention of singly- and multiply-phosphorylated peptides; (4) compatibility with automated, online LC-MS analysis. We demonstrate the reproducibility of RP-SAX-RP and apply it to the analysis of phosphopeptides derived from multiple biological contexts, including an in vitro model of acute myeloid leukemia in addition to primary polyclonal CD8(+) T-cells activated in vivo through bacterial infection and then purified from a single mouse.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.