PUBLICATION
Rhodamine B-conjugated encrypted vipericidin nonapeptide is a potent toxin to zebrafish and associated with in vitro cytotoxicity
- Authors
- Wang, L., Chan, J.Y., Rêgo, J.V., Chong, C.M., Ai, N., Falcão, C.B., Rádis-Baptista, G., Lee, S.M.
- ID
- ZDB-PUB-150304-7
- Date
- 2015
- Source
- Biochimica et biophysica acta. General subjects 1850(6): 1253-60 (Journal)
- Registered Authors
- Keywords
- Rhodamine B-conjugated peptide, cell-penetrating peptide, cytotoxin, encrypted peptides, membrane budding, membranolytic peptide, vipericidins, zebra-fish model
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Antineoplastic Agents/isolation & purification
- Antineoplastic Agents/metabolism
- Antineoplastic Agents/pharmacology*
- Antineoplastic Agents/toxicity
- Breast Neoplasms/metabolism
- Breast Neoplasms/pathology*
- Calcium Signaling/drug effects
- Cathelicidins/isolation & purification
- Cathelicidins/metabolism
- Cathelicidins/pharmacology*
- Cathelicidins/toxicity
- Cell Membrane/drug effects
- Cell Membrane/pathology
- Cell Survival/drug effects
- Dose-Response Relationship, Drug
- Female
- Humans
- Larva/drug effects
- Lethal Dose 50
- MCF-7 Cells
- Molecular Sequence Data
- Oligopeptides/isolation & purification
- Oligopeptides/metabolism
- Oligopeptides/pharmacology*
- Oligopeptides/toxicity
- Rhodamines/metabolism
- Rhodamines/pharmacology*
- Rhodamines/toxicity
- Time Factors
- Viper Venoms/chemistry*
- Zebrafish/embryology*
- PubMed
- 25731980 Full text @ BBA General Subjects
Citation
Wang, L., Chan, J.Y., Rêgo, J.V., Chong, C.M., Ai, N., Falcão, C.B., Rádis-Baptista, G., Lee, S.M. (2015) Rhodamine B-conjugated encrypted vipericidin nonapeptide is a potent toxin to zebrafish and associated with in vitro cytotoxicity. Biochimica et biophysica acta. General subjects. 1850(6):1253-60.
Abstract
Background Animal venoms contain a diverse array of proteins and enzymes that are toxic towards various physiological systems. However, there are also some practical medicinal uses for these toxins including use as anti-bacterial and anti-tumor agents.
Methods in this study, we identified a nine-residue cryptic oligopeptide, KRFKKFFKK (EVP50) that is repeatedly encoded in tandem within vipericidin sequences RESULTS: EVP50 displayed in vivo potent lethal toxicity to zebrafish larvae (LD50 = 6μM) when the peptide's N-terminus was chemically conjugated to rhodamine B (RhoB). In vitro, RhoB-conjugated EVP50 (RhoB-EVP50) exhibited a concentration-dependent cytotoxic effect towards MCF-7 and MDA-MB-231 breast cancer cells. In MCF-7 cells, the RhoB-EVP50 nonapeptide accumulated inside the cells within minutes. In the cytoplasm, the RhoB-EVP50 induced extracellular calcium influx and intracellular calcium release. Membrane budding was also observed after incubation with micromolar concentrations of the fluorescent EVP50 conjugate.
Conclusions The conjugate's interference with calcium homeostasis, its intracellular accumulation and its induced membrane dysfunction (budding and vacuolization) seem to act in concert to disrupt the cell circuitry. Contrastively, unconjugated EVP50 peptide did not display neither toxic nor cytotoxic activities in our in vivo and in vitro models.
General significance The synergic mechanism of toxicity was restricted to the structurally modified encrypted vipericidin nonapeptide.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping