Proteolysis of native proteins. Trapping of a reaction intermediate

Chenyi Wu, Duncan H L Robertson, Simon J. Hubbard, Simon J. Gaskell, Robert J. Beynon

    Research output: Contribution to journalArticlepeer-review


    When limited proteolysis of the mouse major urinary proteins by trypsin was stopped by rapid denaturation of the proteinase, a covalent adduct of the two proteins was observed. The formation of this complex required active trypsin, was favored at low pH, and could be reversed by the addition of covalent or non-covalent trypsin inhibitors. Electrospray mass spectrometry of the complex demonstrated that it was an acyl-enzyme complex, formed after an unusual exopeptidase attack on the C-terminal-Arg-Glu-OH sequence by trypsin. The complex could sequester over 50% of the trypsin in a digestion mixture, and as anticipated, the protein was an effective trypsin inhibitor.
    Original languageEnglish
    Pages (from-to)1108-1115
    Number of pages7
    JournalJournal of Biological Chemistry
    Issue number2
    Publication statusPublished - 8 Jan 1999


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