Abstract
LFchimera, a construct combining two antimicrobial domains of bovine lactoferrin, lactoferrampin265–284 and lactoferricin17–30, possesses strong bactericidal activity. As yet, no experimental evidence was presented to evaluate the mechanisms of LFchimera against Burkholderia isolates. In this study we analyzed the killing activity of LFchimera on the category B pathogen Burkholderia pseudomallei in comparison to the lesser virulent Burkholderia thailandensis often used as a model for the highly virulent B. pseudomallei. Killing kinetics showed that B. thailandensis E264 was more susceptible for LFchimera than B. pseudomallei 1026b. Interestingly the bactericidal activity of LFchimera appeared highly pH dependent; B. thailandensis killing was completely abolished at and below pH 6.4. FITC-labeled LFchimera caused a rapid accumulation within 15 min in the cytoplasm of both bacterial species. Moreover, freeze-fracture electron microscopy demonstrated extreme effects on the membrane morphology of both bacterial species within 1 h of incubation, accompanied by altered membrane permeability monitored as leakage of nucleotides. These data indicate that the mechanism of action of LFchimera is similar for both species and encompasses disruption of the plasma membrane and subsequently leakage of intracellular nucleotides leading to cell dead.
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Acknowledgments
This work was supported by the Commission on Higher Education granting under the CHE-Ph.D.-SW and the Higher Education Research Promotion and National Research University Project of Thailand, Office of the Higher Education Commission, through the Health Cluster (SHeP-GMS), Khon Kaen University. JGMB, KN and ECIV are supported by a grant from the University of Amsterdam for research into the focal point Oral Infections and Inflammation.
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Kanthawong, S., Puknun, A., Bolscher, J.G.M. et al. Membrane-active mechanism of LFchimera against Burkholderia pseudomallei and Burkholderia thailandensis . Biometals 27, 949–956 (2014). https://doi.org/10.1007/s10534-014-9760-5
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DOI: https://doi.org/10.1007/s10534-014-9760-5