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Identification and Characterization of Plasma Kallikrein-Kinin System Inhibitors from Salivary Glands of the Blood-Sucking Insect Triatoma Infestans

Authors: Haruhiko Isawa, Yuki Orito, Naruhiro Jingushi, Siroh Iwanaga, Akihiro Morita, Yasuo Chinzei, Masao Yuda

Field: Medical Entomology, Medical Zoology

Document Content: This study identifies and characterizes two plasma kallikrein-kinin system inhibitors, named triafestin-1 and triafestin-2, from the salivary glands of the kissing bug Triatoma infestans. These inhibitors were found to inhibit the activation of the kallikrein-kinin system by preventing the reciprocal activation of factor XII and prekallikrein, thereby reducing bradykinin release. Binding analyses revealed that triafestin-1 and triafestin-2 specifically interact with factor XII and high molecular weight kininogen in a zinc-dependent manner, suggesting recognition of zinc-induced conformational changes. The inhibitors also interfere with the binding of these proteins to negatively charged surfaces, hindering the initiation of the kallikrein-kinin system. The research highlights the biological significance of kallikrein-kinin system inhibition for blood-feeding arthropods.

Detailed Table of Contents:

  • Keywords
  • Correspondence
  • Abstract
  • Introduction
  • Results
    • cDNA cloning and expression of recombinant triafestin-1 and triafestin-2
    • Triafestin inhibits the plasma kallikrein-kinin system
    • Triafestin inhibits FXII and PK reciprocal activation and bradykinin generation
    • Triafestin binds to both FXII and HK
    • Triafestin inhibits FXII and HK binding to negatively charged surfaces
    • Triafestin interacts with both the N-terminal region of FXII and domain D5 of HK
  • Discussion
  • Experimental procedures
    • Materials
    • Isolation and sequencing of cDNA clones
    • Production and purification of the recombinant proteins
    • Antibody preparation and western blot analysis
    • Expression and purification of the recombinant N-terminal region of FXII and domain D5 of HK
    • Assay for the effect of triafestin-1 and triafestin-2 on plasma coagulation and tenase activity
    • Assay for the effect of triafestin-1 and triafestin-2 on activation of the kallikrein-kinin system
    • Assay for the effect of triafestin-1 and triafestin-2 on bradykinin generation
    • Binding analysis with SPR
  • Acknowledgements
  • References