Beyond the known cuts: trypsin specificity in native proteins

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. © 2025 The Royal Society of Chemistry.

Авторы
Gaspar Marcelo 1, 2, 5 , Sokolova Bohdana 3 , Saei Amir Ata 3, 4 , Marques José Carlos Antunes 2, 5 , Zubarev Roman A. 2, 3, 6, 7
Журнал
Chemical Communications
Издательство
Royal Society of Chemistry
Номер выпуска
68
Язык
Английский
Страницы
12753-12756
Статус
Опубликовано
Ссылка
Внешняя ссылка
DOI
10.1039/D5CC02378E
Том
61
Год
2025
Организации
  • 1 Faculty of Exact Sciences and Engineering, Universidade da Madeira, Funchal, Portugal
  • 2 ISOPlexis Centre Sustainable Agriculture and Food Technology, Universidade da Madeira, Funchal, Portugal
  • 3 Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Stockholms, Sweden
  • 4 Department of Microbiology, Karolinska Institutet, Stockholm, Stockholms, Sweden
  • 5 Nanomodelling and Nanofabrication, Universidade de Aveiro, Aveiro, Aveiro, Portugal
  • 6 Department of Pharmacological & Technological Chemistry, Sechenov First Moscow State Medical University, Moscow, Russian Federation
  • 7 Department of Pharmaceutical and Toxicological Chemistry, RUDN University, Moscow, Moscow Oblast, Russian Federation
Ключевые слова
Genetic engineering; Proteomics; Cleavage preference; Cleavage rates; Context dependent; HeLa cell lysate; Iso-electric points; Native proteins; Protein engineering; Lanthanum compounds; biological marker; elastin-like polypeptide; trypsin; lysine; peptide; protein; amino acid sequence; Article; binding affinity; blood pressure measurement; clinical effectiveness; controlled study; digestion; enzyme activity; hydrolysis; hydrophobicity; immune response; isoelectric focusing; mass spectrometry; nonhuman; optimal digestibility; peptide mapping; physicochemical model; polyacrylamide gel electrophoresis; protein analysis; protein degradation; protein purification; protein synthesis; chemistry; enzyme specificity; HeLa cell line; human; metabolism; HeLa Cells; Humans; Peptides; Proteins; Substrate Specificity
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