Gene cloning and expression of a partial sequence of Hirudomacin, an antimicrobial protein that is increased in leech (Hirudo nipponica Whitman) after a blood meal
Basic science / preclinical published in Comp Biochem Physiol B (2019)
Abstract
The novel antimicrobial gene Hirudomacin (Hmc), with a 249-bp cDNA, encodes a mature protein of 61 amino acids and a 22-amino acid signal peptide. Hmc exhibits the highest similarity, at 90.1%, with macin family members found in the salivary gland of the leech Hirudo nipponica Whitman. A mature Hmc protein concentration of 219 μg/mL was detected using the Bradford method. The mature Hmc protein is 6862.82 Da and contains 8 cysteine residues. Antimicrobial assays showed a minimum bactericidal concentration and 50% lethal dose of 1.56 μg/mL and 0.78 μg/mL, respectively, for Staphylococcus aureus and 0.39 μg/mL and 0.195 μg/mL, respectively, for Bacillus subtilis. Transmission electron microscopy revealed membrane integrity disruption in S. aureus and B. subtilis, which resulted in bacterial lysis. The level of Hmc mRNA in the salivary gland during three blood meal stages indicated a remarkable trend of increase (P < .05), and western blotting demonstrated that among the three blood meal stages, expression of the mature Hmc protein was highest in both the salivary gland and intestine at the fed stage (P < .05). Immunofluorescence further showed the mature Hmc protein to be localized outside the cell nucleus, with the signal intensity in the salivary gland peaking at the fed stage (P < .05). In conclusion, the mature Hmc protein exhibits broad-spectrum antimicrobial effects against gram-positive and gram-negative bacteria, and a blood meal upregulates Hmc gene and protein expression in H. nipponica.
Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.
Zusammenfassung
Cloning and characterization of Hirudomacin (Hmc), a 61-amino-acid antimicrobial macin from Hirudo nipponica. Effective against Staphylococcus aureus (MBC 1.56 ug/mL) and Bacillus subtilis (MBC 0.39 ug/mL); upregulated after blood meal.
Warum dies für die Hirudotherapie relevant ist
Diese Studie klonierte und charakterisierte Hirudomacin (Hmc), ein neuartiges antimikrobielles Protein aus der Speicheldrüse von Hirudo nipponica Whitman, das ein reifes Protein aus 61 Aminosäuren mit 8 Cysteinresten kodiert und breitbandige bakterizide Aktivität gegen S. aureus und B. subtilis durch Membranstörung zeigte. Die Autoren zeigten, dass die Hmc-mRNA- und Proteinexpression in der Speicheldrüse und im Darm nach einer Blutmahlzeit signifikant anstieg und im gefütterten Stadium ihren Höhepunkt erreichte. Dies ist für den ASH-Bereich relevant, da es eine bioaktive antimikrobielle Komponente des Sekretoms des medizinischen Blutegels charakterisiert, obwohl die Befunde präklinisch und auf antimikrobielle In-vitro-Assays sowie Expressionsprofilierung beschränkt sind.
Zitation
Gene cloning and expression of a partial sequence of Hirudomacin, an antimicrobial protein that is increased in leech (Hirudo nipponica Whitman) after a blood meal.
Ding A et al. · Comparative biochemistry and physiology. Part B, 2019
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