ArticleMolecular & cellular proteomics : MCP2011
Pressurized pepsin digestion in proteomics: an automatable alternative to trypsin for integrated top-down bottom-up proteomics.
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 20 papers.
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Who cites it
20 citing papers in PubMed, 51 citations in OpenAlex.
- Optimized Digestion Conditions for Membrane Protein Footprinting and Mass Spectrometry Analysis.Membranes · 2026Article
- Pepsin Digestion for Proteomic Studies of the Human Hair Shaft.Rapid communications in mass spectrometry : RCM · 2026Article
- MSe Collision Energy Optimization for the Analysis of Membrane Proteins Using HDX-cIMS.Journal of the American Society for Mass Spectrometry · 2024Article
- Comparative Label-Free Proteomics Study on Celiac Disease-Active Epitopes in Common Wheat, Spelt, Durum Wheat, Emmer, and Einkorn.Journal of agricultural and food chemistry · 2024Article
- Analysis and Comparison of Proteomics of Placental Proteins from Cows Using Different Proteases.Animals : an open access journal from MDPI · 2023Article
- Targeted Bottom-Up Mass Spectrometry Approach for the Relative Quantification of Post-Translational Modification of Bovine κ-Casein during Milk Fermentation.Molecules (Basel, Switzerland) · 2022Article
- Advances in Hydrogen/Deuterium Exchange Mass Spectrometry and the Pursuit of Challenging Biological Systems.Chemical reviews · 2022Review
- ProAlanase is an Effective Alternative to Trypsin for Proteomics Applications and Disulfide Bond Mapping.Molecular & cellular proteomics : MCP · 2020Article
- Organic Solvents for Enhanced Proteolysis of Stable Proteins for Hydrogen-Deuterium Exchange Mass Spectrometry.Analytical chemistry · 2020Article
- Interlaboratory Comparison of Hydrogen-Deuterium Exchange Mass Spectrometry Measurements of the Fab Fragment of NISTmAb.Analytical chemistry · 2019Article
- Microfluidic-based solid phase extraction of cell free DNA.Lab on a chip · 2018Article
- Quantitative Proteomic Analysis of Mass Limited Tissue Samples for Spatially Resolved Tissue Profiling.Methods in molecular biology (Clifton, N.J.) · 2018Article
- Advances in microscale separations towards nanoproteomics applications.Journal of chromatography. A · 2017Review
- Integral membrane proteins: bottom-up, top-down and structural proteomics.Expert review of proteomics · 2017Review
- Six alternative proteases for mass spectrometry-based proteomics beyond trypsin.Nature protocols · 2016Article
- Accessing the reproducibility and specificity of pepsin and other aspartic proteases.Biochimica et biophysica acta · 2013Article
- Pepsin immobilized on high-strength hybrid particles for continuous flow online digestion at 10,000 psi.Analytical chemistry · 2012Article
- Fully automated multifunctional ultrahigh pressure liquid chromatography system for advanced proteome analyses.Journal of proteome research · 2012Article
- Mass spectrometry-based proteomics: existing capabilities and future directions.Chemical Society reviews · 2012Review
- Uncovering the proteome response of the master circadian clock to light using an AutoProteome system.Molecular & cellular proteomics : MCP · 2011Article
Corrections and comments
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Authors and funding
12 authors at 3 institutions in 2 countries.
Funding
Abstract
Integrated top-down bottom-up proteomics combined with on-line digestion has great potential to improve the characterization of protein isoforms in biological systems and is amendable to high throughput proteomics experiments. Bottom-up proteomics ultimately provides the peptide sequences derived from the tandem MS analyses of peptides after the proteome has been digested. Top-down proteomics conversely entails the MS analyses of intact proteins for more effective characterization of genetic variations and/or post-translational modifications. Herein, we describe recent efforts toward efficient integration of bottom-up and top-down LC-MS-based proteomics strategies. Since most proteomics separations utilize acidic conditions, we exploited the compatibility of pepsin (where the optimal digestion conditions are at low pH) for integration into bottom-up and top-down proteomics work flows. Pressure-enhanced pepsin digestions were successfully performed and characterized with several standard proteins in either an off-line mode using a Barocycler or an on-line mode using a modified high pressure LC system referred to as a fast on-line digestion system (FOLDS). FOLDS was tested using pepsin and a whole microbial proteome, and the results were compared against traditional trypsin digestions on the same platform. Additionally, FOLDS was integrated with a RePlay configuration to demonstrate an ultrarapid integrated bottom-up top-down proteomics strategy using a standard mixture of proteins and a monkey pox virus proteome.
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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.