American Society of Hirudotherapy

Separation of monomerizing and lysozyme activities of destabilase from medicinal leech salivary gland secretion

Basic science published in Biochem Mosc (2001)

Last Updated: June 18, 2026Reviewed by: ASH Editorial Board
Research article — evidence reviewArticle reference
Evidence: Preclinical (animal)Genomics & ProteomicsSalivary PharmacologyBaskova IP et al. · Biochemistry (Mosc), 2001

Abstract

Destabilase, endo-epsilon-(gamma-Glu)-Lys-isopeptidase, was prepared from the salivary gland secretion of the medicinal leech (Hirudo medicinalis). The secretion prepared by the known method of Rigbi et al. (1987) (secretion-K) lacks the destabilase-characteristic highly specific isopeptidase activity (the D-dimer-monomerizing activity) because of its degradation by proteolytic activity (the substrate of Glp-Ala-Ala-Leu-pNA) due to contamination with leech intestinal channel contents. Therefore, we have elaborated a new technique for preparation of a true leech secretion (secretion-I). This secretion is characterized by the complete absence of the leech intestinal channel contents and has no proteolytic activity. For the first time the destabilase-specific D-dimer-monomerizing and lysozyme activities were separated by fractionation of secretion-I by HPLC gel filtration through Superose S-12. For the purified destabilase preparation, these activities were separated by reversed-phase chromatography in an acetonitrile gradient (0-60%) in the presence of 0.1% trifluoroacetic acid. The monomerizing activity of destabilase is responsible for the ability of secretion-I to dissolve stabilized fibrin via isopeptidolysis of alpha-alpha and gamma-gamma fibrin chains bound by epsilon-(gamma-Glu)-Lys-isopeptide bonds.

Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.

Publication typeJournal ArticleResearch Support, Non-U.S. Gov't
Indexed MeSH termsAnimalsEndopeptidasesFibrinLeechesMuramidaseSalivary Glands

Summary

Secretion-I free of intestinal contaminants enables HPLC separation of destabilase's monomerizing (D-dimer cleavage) and lysozyme activities, clarifying dual-functional mechanism.

Why This Matters for Hirudotherapy

This study focused on the biochemical preparation and characterization of destabilase from Hirudo medicinalis salivary gland secretion, addressing problems with contamination by intestinal proteolytic activity in standard preparation methods. The investigators developed a new technique to obtain uncontaminated leech secretion and, for the first time, separated the D-dimer-monomerizing activity (isopeptidase) from lysozyme activity using HPLC gel filtration and reversed-phase chromatography. This work is relevant to understanding the leech secretome's complexity, confirming that destabilase's monomerizing activity dissolves stabilized fibrin through isopeptidolysis of cross-linked fibrin chains. This is a pure biochemistry study with no clinical or in vivo component; its relevance is to the molecular characterization of leech-derived enzymes rather than to therapeutic hirudotherapy outcomes.

Citation

Separation of monomerizing and lysozyme activities of destabilase from medicinal leech salivary gland secretion.

Baskova IP et al. · Biochemistry (Mosc), 2001

DOI

Added to ASH library: May 27, 2026 · Site last updated: June 18, 2026

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