ArticleNature protocols2011
Selecting protein N-terminal peptides by combined fractional diagonal chromatography.
Article in Nature protocols, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 75 papers.
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Who cites it
75 citing papers in PubMed, 178 citations in OpenAlex.
- Mechanisms of Congenital Heart Disease Caused by NAA15 Haploinsufficiency.Circulation research · 2021Trial
- TRAINSPOTTER: profiling nascent protein N-termini indicative of bacterial translation initiation via deformylation-assisted N-terminomics.Nucleic acids research · 2026Article
- Article
- A snapshot of the Physcomitrella N-terminome reveals N-terminal methylation of organellar proteins.Plant cell reports · 2024Article
- Seed longevity is controlled by metacaspases.Nature communications · 2024Article
- Alternative proteoforms and proteoform-dependent assemblies in humans and plants.Molecular systems biology · 2024Article
- An N terminomics toolbox combining 2-pyridinecarboxaldehyde probes and click chemistry for profiling protease specificity.Cell chemical biology · 2024Article
- N-terminal acetylation shields proteins from degradation and promotes age-dependent motility and longevity.Nature communications · 2023Article
- Loss of N-terminal acetyltransferase A activity induces thermally unstable ribosomal proteins and increases their turnover in Saccharomyces cerevisiae.Nature communications · 2023Article
- Structure-function study of a CaProceedings of the National Academy of Sciences of the United States of America · 2023Article
- Proteomic data and structure analysis combined reveal interplay of structural rigidity and flexibility on selectivity of cysteine cathepsins.Communications biology · 2023Article
- Sensitive and High-Throughput Exploration of Protein N-Termini by TMT-TAILS N-Terminomics.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Direct Identification of Proteolytic Cleavages on Living Cells Using a Glycan-Tethered Peptide Ligase.ACS central science · 2022Article
- Protein Processing in Plant Mitochondria Compared to Yeast and Mammals.Frontiers in plant science · 2022Review
- Protein N-terminal acylation: An emerging field in bacterial cell physiology.Current trends in microbiology · 2022Article
- A Strong Cation Exchange Chromatography Protocol for Examining N-Terminal Proteoforms.Methods in molecular biology (Clifton, N.J.) · 2022Article
- The Arabidopsis PeptideAtlas: Harnessing worldwide proteomics data to create a comprehensive community proteomics resource.The Plant cell · 2021Article
- Charting the N-Terminal Acetylome: A Comprehensive Map of Human NatA Substrates.International journal of molecular sciences · 2021Article
- N-Terminal Acetyltransferase Naa40p Whereabouts Put into N-Terminal Proteoform Perspective.International journal of molecular sciences · 2021Article
- Mapping proteolytic neo-N termini at the surface of living cells.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
15 more citing papers are in PubMed but not listed here.
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Authors and funding
9 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In recent years, procedures for selecting the N-terminal peptides of proteins with analysis by mass spectrometry have been established to characterize protease-mediated cleavage and protein α-N-acetylation on a proteomic level. As a pioneering technology, N-terminal combined fractional diagonal chromatography (COFRADIC) has been used in numerous studies in which these protein modifications were investigated. Derivatization of primary amines--which can include stable isotope labeling--occurs before trypsin digestion so that cleavage occurs after arginine residues. Strong cation exchange (SCX) chromatography results in the removal of most of the internal peptides. Diagonal, reversed-phase peptide chromatography, in which the two runs are separated by reaction with 2,4,6-trinitrobenzenesulfonic acid, results in the removal of the C-terminal peptides and remaining internal peptides and the fractionation of the sample. We describe here the fully matured N-terminal COFRADIC protocol as it is currently routinely used, including the most substantial improvements (including treatment with glutamine cyclotransferase and pyroglutamyl aminopeptidase to remove pyroglutamate before SCX, and a sample pooling scheme to reduce the overall number of liquid chromatography-tandem mass spectrometry analyses) that were made since its original publication. Completion of the N-terminal COFRADIC procedure takes ~5 d.
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