Isolation of Novel Peptides, Cabin-1, -2, -3, and -4, That Inhibit Cathepsin B from a Thermolysin Digest Human Plasma

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  • Nakagomi Kazuya
    Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University Faculty of Pharmaceutical Sciences, Teikyo University
  • Fujimura Akiyoshi
    Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University
  • Maeda Hiromi
    Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University
  • Sadakane Yutaka
    Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University
  • Fujii Noriko
    Research Reactor Institute, Kyoto University
  • Akizawa Toshifumi
    Faculty of Pharmaceutical Sciences, Setsunan University
  • Tanimura Takenori
    Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University
  • Hatanaka Yasumaru
    Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University

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  • Isolation of Novel Peptides, Cabin-1, -2, -3, and -4, That Inhibit Cathepsin B from a Thermolysin Digest of Human Plasma.

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Four novel peptides that inhibit cathepsin B, designated as Cabin-1, -2, -3, and -4, were isolated from a thermolysin digest of human plasma. After gel filtration and cation-exchange chromatography, the peptide mixture was purified by reverse-phase HPLC to isolate Cabin-1, -2, -3, and 4, with the amino acid sequences LGPVTQE, VLQSSGLYS, VVSVLT, and LVYDAY, respectively. These peptides correspond to f(64—70) of human apolipoprotein A-I for Cabin-1, f(56—64) and f(185—190) of the human immunoglobulin G γ chain for Cabin-2 and -3, and f(66—71) of human transferrin for Cabin-4. Synthetic Cabin-1, -2, -3, and -4 showed dose-dependent inhibition of cathepsin B. Their IC50 values were 450, 500, 20, and 5.0 μmol/l, respectively. Lineweaver–Burk plots suggested that Cabin-3 is a noncompetitive inhibitor, while Cabin-4 is a competitive inhibitor. Among the N- and C-terminal deletion peptides of Cabin-2 and -4, Cabin-2(1—8), VLQSSGLY, was found to have the most potent inhibitory activity, with an IC50 of 3.8 μmol/l.

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