Defining the importance of the arginine loop region of protegrin-1 for antimicrobial activity towards colistin-resistant Klebsiella pneumoniae

Carbapenem resistant Klebsiella pneumoniae extreme drug resistance has warranted the use of colistin as a last-resort antibiotic but these isolates are now displaying increasing rates of colistin resistance. This is in large part due to the modifications made to the lipid A by the PhoPQ two component system. Host defense peptides (HDPs) are like colistin in that they are both positively charged and display amphipathic character, however are not impacted by lipid A modifications to the extent of colistin. To understand how HDPs can penetrate colistin resistant membranes we performed a deep mutational scanning analysis of protegrin-1 and revealed that amino acids mutations that resulted in alteration of peptide structure had more impact on antimicrobial activity than a reduction in charge. Probing single and double amino acid variants using membrane analysis and molecular modeling revealed the loss of antimicrobial activity correlated with decreased innxser membrane leakage and pore modeling predicting decreased pore size.