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.
Резюме
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.
Почему это важно для гирудотерапии
This study cloned and characterized Hirudomacin (Hmc), a novel antimicrobial protein from the salivary gland of Hirudo nipponica Whitman, encoding a 61-amino acid mature protein with 8 cysteine residues that showed broad-spectrum bactericidal activity against S. aureus and B. subtilis via membrane disruption. The authors demonstrated that Hmc mRNA and protein expression in the salivary gland and intestine increased significantly after a blood meal, peaking at the fed stage. This is relevant to ASH's domain as it characterizes a bioactive antimicrobial component of the medicinal leech secretome, though the findings are preclinical and limited to in vitro antimicrobial assays and expression profiling.
Цитирование
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
Связанный клинический контекст
Узнайте, как это исследование связано с клинической практикой
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