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Beyond the known cuts: trypsin specificity in native proteins

João M. Gaspar, Bohdana Sokolova, Amir Ata Saei, José C. Marques, Roman A. Zubarev

2025Chemical Communications7 citationsDOIOpen Access PDF

Abstract

Above-filter digestion proteomics (AFDIP) was applied to quantify trypsin cleavage preferences in native HeLa cell lysates. Lysine sites were cleaved faster than arginine ones, with cleavage rates modulated by the peptide's size and isoelectric point. These trends, absent in denatured proteomes, highlight trypsin's context-dependent behavior and inform protein engineering for optimal digestibility.

Topics & Concepts

TrypsinCleavage (geology)LysineHeLaArginineChemistryPeptideBiochemistryProteomicsCellBiologyEnzymeAmino acidPaleontologyGeneFracture (geology)Advanced Proteomics Techniques and ApplicationsProtein Structure and DynamicsMachine Learning in Bioinformatics
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