Mechanistic and structural basis of bioengineered bovine Cathelicidin-5 with optimized therapeutic activity.
Peptide-drug discovery using host-defense peptides becomes promising against antibiotic-resistant pathogens and cancer cells. Here, we customized the therapeutic activity of bovine cathelicidin-5 targeting to bacteria, protozoa, and tumor cells. The membrane dependent conformational adaptability and plasticity of cathelicidin-5 is revealed by biophysical analysis and atomistic simulations over 200 μs in thymocytes, leukemia, and E. coli cell-membranes. Our understanding of energy-dependent cathelicidin-5 intrusion in heterogeneous membranes aided in designing novel loss/gain-of-function analogues. In vitro findings identified leucine-zipper to phenylalanine substitution in cathelicidin-5 (1-18) significantly enhance the antimicrobial and anticancer activity with trivial hemolytic activity. Targeted mutants of cathelicidin-5 at kink region and N-terminal truncation revealed loss-of-function. We ensured the existence of a bimodal mechanism of peptide action (membranolytic and non-membranolytic) in vitro. The melanoma mouse model in vivo study further supports the in vitro findings. This is the first structural report on cathelicidin-5 and our findings revealed potent therapeutic application of designed cathelicidin-5 analogues.
Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka, 565-0871, Japan.|Immunology Frontier Research Center, Osaka University, 3-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Bikash R
BR
Sahoo
Immunology Frontier Research Center, Osaka University, 3-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Kenta
K
Maruyama
Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Jyotheeswara R
JR
Edula
Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Takahiro
T
Tougan
Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Yuxi
Y
Lin
Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Young-Ho
YH
Lee
Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Toshihiro
T
Horii
Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Toshimichi
T
Fujiwara
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000032:analysis / Q000737:chemistry* / Q000627:therapeutic use*; D001499:Bayes Theorem; D057005:Bioengineering; D002151:Calorimetry; D002417:Cattle; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D007364:Intercalating Agents / Q000737:chemistry; D008546:Melanoma, Experimental / Q000188:drug therapy / Q000473:pathology; D008563:Membrane Lipids / Q000737:chemistry; D008810:Mice, Inbred C57BL; D008954:Models, Biological; D056004:Molecular Dynamics Simulation; D011485:Protein Binding; D013816:Thermodynamics
D023181:Antimicrobial Cationic Peptides; D007364:Intercalating Agents; D008563:Membrane Lipids; D004247:DNA