TY - JOUR
T1 - Synthetic antimicrobial peptides delocalize membrane bound proteins thereby inducing a cell envelope stress response
AU - Omardien, Soraya
AU - Drijfhout, Jan W.
AU - van Veen, Henk
AU - Schachtschabel, Soraya
AU - Riool, Martijn
AU - Hamoen, Leendert W.
AU - Brul, Stanley
AU - Zaat, Sebastian A.J.
N1 - With supplementary file
PY - 2018/11
Y1 - 2018/11
N2 - Background: Three amphipathic cationic antimicrobial peptides (AMPs) were characterized by determining their effect on Gram-positive bacteria using Bacillus subtilis strain 168 as a model organism. These peptides were TC19 and TC84, derivatives of thrombocidin-1 (TC-1), the major AMPs of human blood platelets, and Bactericidal Peptide 2 (BP2), a synthetic designer peptide based on human bactericidal permeability increasing protein (BPI). Methods: To elucidate the possible mode of action of the AMPs we performed a transcriptomic analysis using microarrays. Physiological analyses were performed using transmission electron microscopy (TEM), fluorescence microscopy and various B. subtilis mutants that produce essential membrane bound proteins fused to green fluorescent protein (GFP). Results: The transcriptome analysis showed that the AMPs induced a cell envelope stress response (cell membrane and cell wall). The cell membrane stress response was confirmed with the physiological observations that TC19, TC84 and BP2 perturb the membrane of B. subtilis. Using B. subtilis mutants, we established that the cell wall stress response is due to the delocalization of essential membrane bound proteins involved in cell wall synthesis. Other essential membrane proteins, involved in cell membrane synthesis and metabolism, were also delocalized due to alterations caused by the AMPs. Conclusions: We showed that peptides TC19, TC84 and BP2 perturb the membrane causing essential proteins to delocalize, thus preventing the possible repair of the cell envelope after the initial interference with the membrane. General significance: These AMPs show potential for eventual clinical application against Gram-positive bacterial cells and merit further application-oriented investigation.
AB - Background: Three amphipathic cationic antimicrobial peptides (AMPs) were characterized by determining their effect on Gram-positive bacteria using Bacillus subtilis strain 168 as a model organism. These peptides were TC19 and TC84, derivatives of thrombocidin-1 (TC-1), the major AMPs of human blood platelets, and Bactericidal Peptide 2 (BP2), a synthetic designer peptide based on human bactericidal permeability increasing protein (BPI). Methods: To elucidate the possible mode of action of the AMPs we performed a transcriptomic analysis using microarrays. Physiological analyses were performed using transmission electron microscopy (TEM), fluorescence microscopy and various B. subtilis mutants that produce essential membrane bound proteins fused to green fluorescent protein (GFP). Results: The transcriptome analysis showed that the AMPs induced a cell envelope stress response (cell membrane and cell wall). The cell membrane stress response was confirmed with the physiological observations that TC19, TC84 and BP2 perturb the membrane of B. subtilis. Using B. subtilis mutants, we established that the cell wall stress response is due to the delocalization of essential membrane bound proteins involved in cell wall synthesis. Other essential membrane proteins, involved in cell membrane synthesis and metabolism, were also delocalized due to alterations caused by the AMPs. Conclusions: We showed that peptides TC19, TC84 and BP2 perturb the membrane causing essential proteins to delocalize, thus preventing the possible repair of the cell envelope after the initial interference with the membrane. General significance: These AMPs show potential for eventual clinical application against Gram-positive bacterial cells and merit further application-oriented investigation.
KW - Amphipathic peptides
KW - Antimicrobial Cationic Peptides/chemistry
KW - BPI derivative
KW - Bacillus subtilis/drug effects
KW - Bacterial Proteins/genetics
KW - Cell Wall/chemistry
KW - Cell envelope stress
KW - Cluster Analysis
KW - Humans
KW - Membrane Proteins/genetics
KW - Membrane bound proteins
KW - Membrane perturbation
KW - Microbial Sensitivity Tests
KW - Microscopy, Electron, Transmission
KW - Thrombocidin-1 derivatives
KW - Transcriptome/drug effects
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UR - https://www.ncbi.nlm.nih.gov/pubmed/29894683
UR - https://pure.uva.nl/ws/files/34867333/1_s2.0_S0005273618301779_mmc1.docx
U2 - https://doi.org/10.1016/j.bbamem.2018.06.005
DO - https://doi.org/10.1016/j.bbamem.2018.06.005
M3 - Article
C2 - 29894683
SN - 0005-2736
VL - 1860
SP - 2416
EP - 2427
JO - Biochimica et Biophysica Acta - Biomembranes
JF - Biochimica et Biophysica Acta - Biomembranes
IS - 11
ER -