Cloning and Functional Analysis of Skin Host Defense Peptides from Yakushima Tago's Brown Frog (Rana tagoi yakushimensis) and Development of Serum Endotoxin Detection System.

Aono, Taichi; Tamura, Saki; Suzuki, Yua; Imanara, Taichi; Niwa, Ryosei; Yamane, Yoshie; Kobayashi, Tetsuya; Kikuyama, Sakae; Hasunuma, Itaru; Iwamuro, Shawichi

Background/Objective: Amphibian skin is a valuable source of host defense peptides (HDPs). This study aimed to identify HDPs with novel amino acid sequences from the skin of Rana tagoi yakushimensis and analyze their functions. Methods: cDNAs encoding HDP precursors were cloned and sequenced using RT-PCR and 3′-RACE. The novel HDPs were synthesized to evaluate their antimicrobial activity, antioxidant activity, and cytotoxicity. Antimicrobial activity was evaluated by way of broth microdilution and endotoxin- and β-glucan-binding capacity using an enzyme-linked endotoxin binding assay (ELEBA) and a modified ELEBA, respectively. Results: Nine cDNAs encoding precursors for various HDP families, including temporin, ranatuerin-2, brevinin-1, amurin-9, and a novel yakushimin peptide, were identified. Brevinin-1TYa exhibited antibacterial activity against Staphylococcus aureus, and brevinin-1TYa and amurin-9TYa induced morphological changes in Escherichia coli and S. aureus. Yakushimin-TYa, amurin-9TYa, and brevinin-1TYa showed concentration-dependent antibacterial effects against the plant pathogens Xanthomonas oryzae pv. oryzae and Clavibacter michiganensis subsp. michiganensis. Amurin-9TYa demonstrated strong binding affinity to lipopolysaccharide, lipoteichoic acid, and β-glucan, exhibited antioxidant activity, and lacked cytotoxicity, making it a promising therapeutic candidate. Moreover, brevinin-1TYa showed strong cytotoxicity, whereas yakushimin-TYa exhibited weak cytotoxicity. Conclusions: These findings highlight the potential of these peptides, particularly amurin-9TYa, for future applications as antimicrobial and therapeutic agents.

ANTIMICROBIAL peptides; RANA temporaria; CYTOTOXINS; PHYTOPATHOGENIC microorganisms; AMINO acid sequence

Antibiotics (2079-6382), 2024, Vol 13, Issue 12, p1127

2079-6382

Academic Journal

10.3390/antibiotics13121127