ArticleNature protocols2011
Characterization of the prime and non-prime active site specificities of proteases by proteome-derived peptide libraries and tandem mass spectrometry.
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 46 papers.
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Who cites it
46 citing papers in PubMed, 108 citations in OpenAlex.
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- Optimization of quenched fluorescent peptide substrates of SARS-CoV-2 3CLJournal of virology · 2024Article
- An N terminomics toolbox combining 2-pyridinecarboxaldehyde probes and click chemistry for profiling protease specificity.Cell chemical biology · 2024Article
- Mouse skin peptidomic analysis of the hemorrhage induced by a snake venom metalloprotease.Amino acids · 2023Article
- Proteome-Derived Peptide Libraries for Deep Specificity Profiling of N-terminal Modification Reagents.Current protocols · 2023Article
- Making the cut with protease engineering.Cell chemical biology · 2022Review
- Proteolytic Profiling of Streptococcal Pyrogenic Exotoxin B (SpeB) by Complementary HPLC-MS Approaches.International journal of molecular sciences · 2021Article
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- Innate immune evasion mediated by picornaviral 3C protease: Possible lessons for coronaviral 3C-like protease?Reviews in medical virology · 2021Review
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- Identification and characterization of the proteolytic flagellin from the common freshwater bacterium Hylemonella gracilis.Scientific reports · 2020Article
- Structural and functional studies ofThe Journal of biological chemistry · 2020Article
- Sirtilins - the new old members of the vitamin K-dependent coagulation factor family.Journal of thrombosis and haemostasis : JTH · 2019Article
- Ten Prominent Host Proteases in Plant-Pathogen Interactions.International journal of molecular sciences · 2018Review
- Block-based characterization of protease specificity from substrate sequence profile.BMC bioinformatics · 2017Article
- Discovery of a proteolytic flagellin family in diverse bacterial phyla that assembles enzymatically active flagella.Nature communications · 2017Article
Corrections and comments
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Authors and funding
5 authors at 3 institutions in 4 countries.
Funding
Abstract
To link cleaved substrates in complex systems with a specific protease, the protease active site specificity is required. Proteomic identification of cleavage sites (PICS) simultaneously determines both the prime- and non-prime-side specificities of individual proteases through identification of hundreds of individual cleavage sequences from biologically relevant, proteome-derived peptide libraries. PICS also identifies subsite cooperativity. To generate PICS peptide libraries, cellular proteomes are digested with a specific protease such as trypsin. Following protease inactivation, primary amines are protected. After incubation with a test protease, each prime-side cleavage fragment has a free newly formed N-terminus, which is biotinylated for affinity isolation and identification by liquid chromatography-tandem mass spectrometry. The corresponding non-prime sequences are derived bioinformatically. The step-by-step protocol also presents a web service for PICS data analysis, as well as introducing and validating PICS peptide libraries made from Escherichia coli.
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Registered trials
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.