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Comparative Study of the Action of Bovine Duodenal Proteinases (Duodenases) on Polypeptide Substrates

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Abstract

A comparative study of substrate specificity of bovine duodenal proteinases—chymotrypsin-like duodenase (ChlD) and dual-specificity duodenase (dsD)—was carried out using oligopeptide substrates (human proinsulin, glucagon, melittin, angiotensinogen fragment 1-14). ChlD displayed mainly chymotrypsin-like properties towards these substrates, hydrolyzing peptide bonds carboxy-terminally to bulky aliphatic or aromatic residues. In melittin, ChlD additionally cleaved peptide bonds after Thr and Ser residues. Dual-specificity duodenase (dsD) significantly restricted its specificity to only trypsin-like or only chymotrypsin-like or displayed full activity, combining both specificities, depending on substrate. Both ChlD and dsD efficiently hydrolyzed a single peptide bond (Phe8–His9) in angiotensinogen fragment 1-14. The kinetic parameters of angiotensinogen fragment 1-14 cleavage by ChlD and dsD were determined (k cat/K m = 80,500 M-1·sec-1 for ChlD and 103,000 M-1·sec-1 for dsD).

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Authors and Affiliations

  1. Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul, Miklukho-Maklaya 16/10, Moscow, 117871, Russia

    E. A. Sokolova & T. S. Zamolodchikova

  2. Institute of Cytology, Russian Academy of Sciences, Tikhoretskaya nab. 4, St. Petersburg, 194064, Russia

    O. A. Mirgorodskaya & N. V. Savelyeva

  3. Department of Molecular Biology, Odense University, DK-5230, Odense M, Denmark

    P. Roepstorff

Authors
  1. E. A. Sokolova
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  2. O. A. Mirgorodskaya
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  3. P. Roepstorff
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  4. N. V. Savelyeva
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  5. T. S. Zamolodchikova
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Sokolova, E.A., Mirgorodskaya, O.A., Roepstorff, P. et al. Comparative Study of the Action of Bovine Duodenal Proteinases (Duodenases) on Polypeptide Substrates. Biochemistry (Moscow) 66, 62–67 (2001). https://doi.org/10.1023/A:1002833729744

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