Molecular cloning and functional analysis of a destabilase from Hirudinaria manillensis
Basic science / preclinical published in Protein Expr Purif (2025)
Abstract
Destabilases are i-type lysozymes with isopeptidase activity and antibacterial and thrombolytic functions. In recent years, destabliases have been identified in an increasing number of invertebrates. Hirudinaria manillensis belonging to the Annelida, as one of the origins of leeches used in traditional Chinese medicine, which has high medicinal value, there have been few reports on the H. manillensis destabliase. In this study, the cDNA sequence of Hmdestabilase was cloned from the salivary glands of H. manillensis. The 3D Structural analysis indicated that Hmdestabilase is similar to other i-type lysozymes in that it adopts an ellipsoidal shape and has a large cleft containing the lysozyme active site. The docking results of Hmdestabilase protein with N-acetylglucosamine trimer molecule have shown that the location and number of hydrogen bonds are one of the key factors for the interaction between the protein and its substrate. The Hmdestabilase fusion protein obtained through the prokaryotic expression system has lysozyme and isopeptidase activities. In addition, Changes in sodium ion concentration in the environment affect the lysozyme activity of Hmdestabilase fusion protein. The above bioinformatic analysis and enzymatic function studies have shown that Hmdestabilase belongs to the i-type lysozyme family. qPCR analysis revealed that blood feeding significantly increased the mRNA expression of Hmdestabilase in the salivary glands of H. manillensis,and successfully priming the innate immune system against harmful microorganisms ingested with food. This study is helpful to elucidate the innate immune response of H. manillensis and promote the artificial breeding of H. manillensis.
Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.
Zusammenfassung
Cloning and characterization of Hmdestabilase from Hirudinaria manillensis salivary glands. Recombinant protein shows lysozyme and isopeptidase activities; expression upregulated after blood feeding. Demonstrates innate-immunity priming role.
Warum dies für die Hirudotherapie relevant ist
Diese Studie klonierte und charakterisierte Hmdestabilase aus den Speicheldrüsen von Hirudinaria manillensis, einem in der traditionellen chinesischen Medizin verwendeten medizinischen Blutegel. Das rekombinante Fusionsprotein zeigte Lysozym- und Isopeptidase-Aktivitäten, und das Blutsaugen führte zu einer Hochregulierung der mRNA-Expression in den Speicheldrüsen, was mit Rollen in der angeborenen Immunität und Thrombolyse vereinbar ist. Für die ASH ist dies relevant, da Destabilasen eine Enzymklasse des Blutegelsekretoms darstellen, die mit potenziellen thrombolytischen und antimikrobiellen Eigenschaften für die Hirudotherapie von Interesse sind. Die Arbeit beschränkt sich auf molekulares Klonen, Bioinformatik und In-vitro-Enzymassays; es werden keine In-vivo-Daten zur Thrombolyse oder therapeutischen Wirksamkeit vorgelegt, und die Befunde belegen keine klinische Wirksamkeit.
Zitation
Molecular cloning and functional analysis of a destabilase from Hirudinaria manillensis.
Gao T et al. · Protein expression and purification, 2025
Verwandter klinischer Kontext
Erfahren Sie, wie diese Forschung mit der klinischen Praxis verknüpft ist
Zur ASH-Bibliothek hinzugefügt: May 27, 2026 · Letzte Aktualisierung der Website: 18. Juni 2026