Rational design, structural characterisation, and functional evaluation of antimicrobial peptides from Rana palustris

This project investigates skin antimicrobial peptides (AMPs) from the frog, Rana palustris, across three families: Temporin, Ranatuerin-2, and Palustrin-1, using an integrated workflow of discovery/confirmation, rational design, structural characterisation, and functional validation.<br><br>A new temporin member, Temporin-1PLb, was cloned and identified, and analogue libraries were generated through targeted substitutions, D-residue incorporation, truncation and motif grafting.

Ranatuerin-2PLb and Palustrin-1c were further used as tunable scaffolds by editing cation density, hydrophobicity, chirality and the rana-box region. All peptides were verified by MS and RP-HPLC, and CD was used to track conformational changes in water and membrane-mimicking conditions.<br><br>Activities and mechanisms were assessed using MIC/MBC, MBIC/MBEC, time-kill kinetics, salt/serum interference, haemolysis and therapeutic index, plus membrane depolarisation, permeability staining and SYTO9/PI imaging.