title: Cell-permeable gomesin peptide promotes cell death by intracellular Ca(2+) overload.
abstract: In recent years, the antitumoral activity of antimicrobial peptides (AMPs) has been the goal of many research studies. Among AMPs, gomesin (Gm) displays antitumor activity by unknown mechanisms. Herein, we studied the cytotoxicity of Gm in the Chinese hamster ovary (CHO) cell line. Furthermore, we investigated the temporal ordering of organelle changes and the dynamics of Ca(2+) signaling during Gm-induced cell death. The results indicated that Gm binds to the plasma membrane and rapidly translocates into the cytoplasm. Moreover, 20 μM Gm increases the cytosolic Ca(2+) and induces  membrane permeabilization after 30 min of treatment. Direct Ca(2+) measurements in CHO cells transfected with the genetically encoded D1-cameleon to the endoplasmic reticulum (ER) revealed that Gm induces ER Ca(2+) depletion, which in turn resulted in oscillatory mitochondrial Ca(2+) signal, as measured in cells expressing the genetically encoded probe to the mitochondrial matrix (mit)Pericam. This leads to mitochondria disruption, loss of mitochondrial membrane potential and increased reactive oxygen species prior to membrane permeabilization. Gm-induced membrane permeabilization by a Ca(2+)-dependent pathway involving Gm translocation into the cell, ER Ca(2+) depletion and disruption, mitochondrial Ca(2+) overload and oxidative stress.
authors.affiliation: Departamento de Bioquímica, Universidade Federal de São Paulo, Três de Maio 100, 04044-020, São Paulo, Brazil.
authors.forename: Edgar J
authors.initials: EJ
authors.lastname: Paredes-Gamero
authors.forename: Rafael L
authors.initials: RL
authors.lastname: Casaes-Rodrigues
authors.forename: Gioconda E D D
authors.initials: GE
authors.lastname: Moura
authors.forename: Tatiana M
authors.initials: TM
authors.lastname: Domingues
authors.forename: Marcus V
authors.initials: MV
authors.lastname: Buri
authors.forename: Victor H C
authors.initials: VH
authors.lastname: Ferreira
authors.forename: Edvaldo S
authors.initials: ES
authors.lastname: Trindade
authors.forename: Ana J
authors.initials: AJ
authors.lastname: Moreno-Ortega
authors.forename: María F
authors.initials: MF
authors.lastname: Cano-Abad
authors.forename: Helena B
authors.initials: HB
authors.lastname: Nader
authors.forename: Alice T
authors.initials: AT
authors.lastname: Ferreira
authors.forename: Antonio
authors.initials: A
authors.lastname: Miranda
authors.forename: Giselle Z
authors.initials: GZ
authors.lastname: Justo
authors.forename: Ivarne L S
authors.initials: IL
authors.lastname: Tersariol
mesh_terms: D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000494:pharmacology*; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism*; D016923:Cell Death / Q000187:drug effects*; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects*; D006224:Cricetinae; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D023181:Antimicrobial Cationic Peptides; D017382:Reactive Oxygen Species; C415987:gomesin; D002118:Calcium