Antimicrobial protegrin-1 forms ion channels: molecular dynamic simulation, atomic force microscopy, and electrical conductance studies.
Antimicrobial peptides (AMPs) are an emerging class of antibiotics for controlling health effects of antibiotic-resistant microbial strains. Protegrin-1 (PG-1) is a model antibiotic among beta-sheet AMPs. Antibiotic activity of AMPs involves cell membrane damage, yet their membrane interactions, their 3D membrane-associated structures and the mechanism underlying their ability to disrupt cell membrane are poorly understood. Using complementary approaches, including molecular dynamics simulations, atomic force microscopy (AFM) imaging, and planar lipid bilayer reconstitution, we provide computational and experimental evidence that PG-1, a beta-hairpin peptide, forms ion channels. Simulations indicate that PG-1 forms channel-like structures with loosely attached subunits when reconstituted in anionic lipid bilayers. AFM images show the presence of channel-like structures when PG-1 is reconstituted in dioleoylphosphatidylserine/palmitoyloleoyl phosphatidylethanolamine bilayers or added to preformed bilayers. Planar lipid bilayer electrical recordings show multiple single channel conductances that are consistent with the heterogeneous oligomeric channel structures seen in AFM images. PG-1 channel formation seems to be lipid-dependent: PG-1 does not easily show ion channel electrical activity in phosphatidylcholine membranes, but readily shows channel activity in membranes rich in phosphatidylethanolamine or phosphatidylserine. The combined results support a model wherein the beta-hairpin PG-1 peptide acts as an antibiotic by altering cell ionic homeostasis through ion channel formation in cell membranes.
Center for Nanomedicine and Department of Medicine, University of Chicago, Chicago, Illinois, USA.
Ricardo
R
Capone
Mirela
M
Mustata
Hyunbum
H
Jang
Fernando Teran
FT
Arce
Ruth
R
Nussinov
Ratnesh
R
Lal
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry*; D004553:Electric Conductivity; D008051:Lipid Bilayers / Q000737:chemistry; D008564:Membrane Potentials; D018625:Microscopy, Atomic Force; D056004:Molecular Dynamics Simulation; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010718:Phosphatidylserines / Q000737:chemistry; D011336:Probability; D017433:Protein Structure, Secondary; D013552:Swine
D023181:Antimicrobial Cationic Peptides; D008051:Lipid Bilayers; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; C094880:protegrin-1; C057561:1-palmitoyl-2-oleoylphosphatidylethanolamine; C042511:1,2-dioleoylphosphatidylserine