Membrane-active epithelial keratin 6A fragments (KAMPs) are unique human antimicrobial peptides with a non-αβ structure

Judy T.Y. Lee, Guangshun Wang, Yu Tong Tam, Connie Tam

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Antibiotic resistance is a pressing global health problem that threatens millions of lives each year. Natural antimicrobial peptides and their synthetic derivatives, including peptoids and peptidomimetics, are promising candidates as novel antibiotics. Recently, the C-terminal glycine-rich fragments of human epithelial keratin 6A were found to have bactericidal and cytoprotective activities. Here, we used an improved 2-dimensional NMR method coupled with a new protocol for structural refinement by low temperature simulated annealing to characterize the solution structure of these kerain-derived antimicrobial peptides (KAMPs). Two specific KAMPs in complex with membrane mimicking sodium dodecyl sulfate (SDS) micelles displayed amphipathic conformations with only local bends and turns, and a central 10-residue glycine-rich hydrophobic strip that is central to bactericidal activity. To our knowledge, this is the first report of non-αβ structure for human antimicrobial peptides. Direct observation of Staphylococcus aureus and Pseudomonas aeruginosa by scanning and transmission electron microscopy showed that KAMPs deformed bacterial cell envelopes and induced pore formation. Notably, in competitive binding experiments, KAMPs demonstrated binding affinities to LPS and LTA that did not correlate with their bactericidal activities, suggesting peptide-LPS and peptide-LTA interactions are less important in their mechanisms of action. Moreover, immunoprecipitation of KAMPs-bacterial factor complexes indicated that membrane surface lipoprotein SlyB and intracellular machineries NQR sodium pump and ribosomes are potential molecular targets for the peptides. Results of this study improve our understanding of the bactericidal function of epithelial cytokeratin fragments, and highlight an unexplored class of human antimicrobial peptides, which may serve as non-αβ peptide scaffolds for the design of novel peptide-based antibiotics.

Original languageEnglish (US)
Article number1799
JournalFrontiers in Microbiology
Volume7
Issue numberNOV
DOIs
StatePublished - Nov 11 2016

Fingerprint

Keratin-6
Peptides
Membranes
Glycine
Peptoids
Anti-Bacterial Agents
Peptidomimetics
Sodium-Potassium-Exchanging ATPase
Scanning Transmission Electron Microscopy
Competitive Binding
Micelles
Microbial Drug Resistance
Keratins
Ribosomes
Immunoprecipitation

Keywords

  • Antimicrobial peptides
  • Epithelial cells
  • Innate immunity
  • Keratins
  • Peptide structure and function

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

Cite this

Membrane-active epithelial keratin 6A fragments (KAMPs) are unique human antimicrobial peptides with a non-αβ structure. / Lee, Judy T.Y.; Wang, Guangshun; Tam, Yu Tong; Tam, Connie.

In: Frontiers in Microbiology, Vol. 7, No. NOV, 1799, 11.11.2016.

Research output: Contribution to journalArticle

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