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Título: | The induction of NOS2 expression by the hybrid cecropin A-melittin antibiotic peptide CA(1-8)M(1-18) in the monocytic line RAW 264.7 is triggered by a temporary and reversible plasma membrane permeation |
Autor: | Arias, Cristina CSIC; Guizy, Miriam CSIC; Luque-Ortega, Juan Román CSIC ORCID ; Guerrero, Esther; García de la Torre, Beatriz CSIC ORCID; Andreu, David; Rivas, Luis CSIC ORCID ; Valenzuela, Carmen CSIC ORCID CVN | Fecha de publicación: | 2006 | Editor: | Elsevier | Citación: | BBA - Molecular Cell Research 1763(1): 110-119 (2006) | Resumen: | There is an increasing awareness of immune cell modulation by antimicrobial peptides. While this process often requires specific receptors for the peptides involved, several reports point out to a receptor-independent process. The cecropin A-melittin hybrid peptide CA(1-8)M(1-18) (KWKLFKKIGIGAVLKVLTTGLPALIS- amide) modifies gene expression in the macrophage line RAW 264.7 in the absence of any previous macrophage priming, suggesting a membrane permeation process. To further analyze the initial steps of this mechanism, we have studied the interaction of the peptide with these cells. Below 2 μM, CA(1-8)M(1-18) causes a concentration-dependent membrane depolarization partially reversible with time. At 2 μM, the accumulation of the SYTOX green vital dye is one half of that achieved with 0.05% Triton X-100. The binding level, as assessed by fluorescein-labeled CA(1-8)M(1-18), varies from 7.7 ± 1.2 to 37.4 ± 3.9 × 106 molecules/cell over a 0.5-4.0 μM concentration range. Electrophysiological experiments with 0.5 μM CA(1-8)M(1-18), a concentration that triggers maximal NOS2 expression and minimal toxicity, show a reversible current induction in the RAW 264.7 plasma membrane that is maintained as far as peptide is present. This activation of the macrophage involves the production of nitric oxide, a metabolite lethal for many pathogens that results from unspecific membrane permeation by antimicrobial peptides, and represents a new mode of action that may open new therapeutic possibilities for these compounds against intracellular pathogens. © 2005 Elsevier B.V. All rights reserved. | URI: | http://hdl.handle.net/10261/81314 | DOI: | 10.1016/j.bbamcr.2005.11.003 | Identificadores: | doi: 10.1016/j.bbamcr.2005.11.003 issn: 0167-4889 |
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