Characterization of dual effects induced by antimicrobial peptides: Regulated cell death or membrane disruption

Author(s)

Paredes-Gamero, Edgar J. ^{[1, 2]} ; Martins, Marta N. C. ^[2] ; Cappabianco, Fabio A. M. ^[3] ; Ide, Jaime S. ^[3] ; Miranda, Antonio ^[2]

Affiliation

^[1] Univ Fed Sao Paulo, Dept Bioquim, BR-04044020 Sao Paulo - Brazil

^[2] Univ Fed Sao Paulo, Dept Biofis, BR-04044020 Sao Paulo - Brazil

^[3] Univ Fed Sao Paulo, Dept Ciencia & Tecnol, BR-12231280 Sao Jose Dos Campos, SP - Brazil

• Document type: Journal article
• Source: BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS; v. 1820, n. 7, p. 1062-1072, JUL 2012.
• Web of Science Citations: 60

Abstract

Background: Some reports describe lysis mechanisms by antimicrobial peptides (AMPs), while others describe the activation of regulated cell death. In this study, we compare the cell death-inducing activities of four beta-hairpin AMPs (gomesin, protegrin, tachyplesin and polyphemusin II) along with their linear analogs in the human erythroleukemia K562 cell line to investigate the relationship between their structure and activity. Methods: K562 cells were exposed to AMPs. Morphological and biochemistry alterations were evaluated using light microscopy, confocal microscopy and flow cytometry. Results: Gomesin and protegrin displayed cytotoxic properties that their linear counterparts did not. Tachyplesin and polyphemusin II and also their linear analogs induced cell death. We were able to distinguish two ways in which these AMPs induced cell death. Lower concentrations of AMPs induced controlled cell death mechanisms. Gomesin, tachyplesin and linear-tachyplesin promoted apoptosis that was characterized by annexin labeling, sensitivity to Z-VAD, and caspase-3 activation, but was also inhibited by necrostatin-1. Gomesin and protegrin induced cell death was dependent on intracellular Ca2+ mechanisms and the participation of free radicals was observed in protegrin induced cell death. Polyphemusin II and its linear analog mainly induced necrosis. Conversely, treatment with higher concentrations of AMPs primarily resulted in cell membrane disruption, but with clearly different patterns of action for each AMP tested. Conclusion: Different actions by beta-hairpin AMPs were observed at low concentrations and at higher concentrations despite the structure similarity. General significance: Controlled intracellular mechanism and direct membrane disruption were clearly distinguished helping to understand the real action of AMPs in mammalian cells. (C) 2012 Elsevier B.V. All rights reserved. (AU)

• FAPESP's process: 11/17584-0 - Studies of the structure-activity relationship and mechanism of lytic action of the antimicrobial peptide gomesin
• Grantee: Antonio de Miranda
• Support Opportunities: Regular Research Grants