ArticleJournal of the American Society for Mass Spectrometry2009
A robust two-dimensional separation for top-down tandem mass spectrometry of the low-mass proteome.
Article in Journal of the American Society for Mass Spectrometry, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 50 papers.
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Who cites it
50 citing papers in PubMed.
- A Draft Map of E. coli Proteoforms.Analytical chemistry · 2026Article
- Proteomics-The State of the Field: The Definition and Analysis of Proteomes Should Be Based in Reality, Not Convenience.Proteomes · 2024Article
- Top-Down Proteomics and the Challenges of True Proteoform Characterization.Journal of proteome research · 2023Review
- Multidimensional Separations in Top-Down Proteomics.Analytical science advances · 2023Article
- TopPICR: A Companion R Package for Top-Down Proteomics Data Analysis.Journal of proteome research · 2023Article
- Mapping the Proteoform Landscape of Five Human Tissues.Journal of proteome research · 2022Article
- Fourier-transform ion cyclotron resonance mass spectrometry for characterizing proteoforms.Mass spectrometry reviews · 2022Review
- RPLC-RPLC-MS/MS for Proteoform Identification.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Development of Nascent Focal Adhesions in Spreading Cells.Biophysical journal · 2020Review
- High-throughput quantitative top-down proteomics.Molecular omics · 2020Review
- Review
- VisioProt-MS: interactive 2D maps from intact protein mass spectrometry.Bioinformatics (Oxford, England) · 2019Article
- Top-Down Protein Identification using a Time-of-Flight Mass Spectrometer and Data Independent Acquisition.International journal of mass spectrometry · 2019Article
- High-Throughput Analysis of Intact Human Proteins Using UVPD and HCD on an Orbitrap Mass Spectrometer.Journal of proteome research · 2017Article
- Elucidating Proteoform Families from Proteoform Intact-Mass and Lysine-Count Measurements.Journal of proteome research · 2016Article
- An Integrated Multiomics Approach to Identify Candidate Antigens for Serodiagnosis of Human Onchocerciasis.Molecular & cellular proteomics : MCP · 2015Article
- Proteomic identification of nuclear processes manipulated by cytomegalovirus early during infection.Proteomics · 2015Article
- Online matrix removal platform for coupling gel-based separations to whole protein electrospray ionization mass spectrometry.Journal of proteome research · 2015Article
- In-line separation by capillary electrophoresis prior to analysis by top-down mass spectrometry enables sensitive characterization of protein complexes.Journal of proteome research · 2014Article
- Sheathless capillary electrophoresis-tandem mass spectrometry for top-down characterization of Pyrococcus furiosus proteins on a proteome scale.Analytical chemistry · 2014Article
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12 authors.
Funding
Abstract
For fractionation of intact proteins by molecular weight (MW), a sharply improved two-dimensional (2D) separation is presented to drive reproducible and robust fractionation before top-down mass spectrometry of complex mixtures. The "GELFrEE" (i.e., gel-eluted liquid fraction entrapment electrophoresis) approach is implemented by use of Tris-glycine and Tris-tricine gel systems applied to human cytosolic and nuclear extracts from HeLa S3 cells, to achieve a MW-based fractionation of proteins from 5 to >100 kDa in 1 h. For top-down tandem mass spectroscopy (MS/MS) of the low-mass proteome (5-25 kDa), between 5 and 8 gel-elution (GE) fractions are sampled by nanocapillary-LC-MS/MS with 12 or 14.5 tesla Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. Single injections give about 40 detectable proteins, about half of which yield automated ProSight identifications. Reproducibility metrics of the system are presented, along with comparative analysis of protein targets in mitotic versus asynchronous cells. We forward this basic 2D approach to facilitate wider implementation of top-down mass spectrometry and a variety of other protein separation and/or characterization approaches.
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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.