Melittin: a Membrane-active Peptide with Diverse Functions

Raghuraman, H.; Chattopadhyay, Amitabha

doi:10.1007/s10540-006-9030-z

Cited by 539 publications

(529 citation statements)

References 267 publications

(428 reference statements)

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“…Apoptosis was not detected in intestinal goblet cells by four methods for detecting programmed cell death [15], but did however detect necrosis by annexin-FITC/PI dual staining. There are multiple studies showing different effects of melittin in different cell types (reviewed in [14,38]). What is clear is that the actions of micromolar concentrations of melittin are mediated, in part, through increases in IC, decreased MMP and loss of membrane integrity, which may lead to concentration-, time-and cell typedependent induction of cell death.…”

Section: Discussionmentioning

confidence: 99%

“…For example, structure activity studies on the AMP, melittin, have generated large numbers of structural analogues [11]. Melittin is an amphipathic cationic linear 26-mer α-peptide that has cell penetrating activity as well as well-known cytotoxicity in erythrocytes [12] and enterocytes [13] and other mammalian cells [14]. Alongside phospholipase A2, it is a major constituent of the venom of Apis Mellifera, and has been found to cause apoptosis and necrosis in different cells [15,16], as well as agonist and antagonist actions on many cellular proteins including phospholipase A2 and ATPases (reviewed in [14]).…”

Section: Introductionmentioning

confidence: 99%

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High content analysis to determine cytotoxicity of the antimicrobial peptide, melittin and selected structural analogs

et al. 2011

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Antimicrobial peptides (AMPs) are naturally occurring entities with potential as pharmaceutical candidates and/or food additives. They are present in many organisms including bacteria, insects, fish and mammals. While their antimicrobial activity is equipotent with many commercial antibiotics, current limitations are poor pharmacokinetics, stability and potential toxicology issues. Most elicit antimicrobial action via perturbation of bacterial membranes. Consequently, associated cytotoxicity in human cells is reflected by their capacity to lyse erythrocytes. However, more rigorous toxicological assessment of AMPs is required in order to predict potential failure at a later stage of development. We describe a high-content analysis (HCA) screening protocol recently established for determination and prediction of safety in pharmaceutical drug discovery. HCA is a powerful, multi-parameter bioanalytical tool that amalgamates the actions of fluorescence microscopy with automated cell analysis software in order to understand multiple changes in cellular health. We describe the application of HCA in assessing cytotoxicity of the cytolytic α-helical peptide, melittin, and selected structural analogues. The data shows that structural modification of melittin reduces its cytotoxic action and that HCA is suitable for rapidly identifying cytotoxicity.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

High content analysis to determine cytotoxicity of the antimicrobial peptide, melittin and selected structural analogs

et al. 2011

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“…As an example, we investigated the effect of the peptide melittin from bee venom, 40 which at micromolar concentrations forms large pores in lipid bilayers, eventually leading to membrane disintegration in a detergent-like fashion. 41 We contacted a 3 μL droplet containing 5 μg mL −1 (≈2 μM) melittin with a linear array of densely packed 1 μL droplets of buffer solution (Fig.…”

Section: Bilayer Permeation and Perturbationmentioning

confidence: 99%

Interdroplet bilayer arrays in millifluidic droplet traps from 3D-printed moulds

et al. 2014

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In droplet microfluidics, aqueous droplets are typically separated by an oil phase to ensure containment of molecules in individual droplets of nano-to-picoliter volume. An interesting variation of this method involves bringing two phospholipid-coated droplets into contact to form a lipid bilayer in-between the droplets. These interdroplet bilayers, created by manual pipetting of microliter droplets, have proved advantageous for the study of membrane transport phenomena, including ion channel electrophysiology.In this study, we adapted the droplet microfluidics methodology to achieve automated formation of interdroplet lipid bilayer arrays. We developed a 'millifluidic' chip for microliter droplet generation and droplet packing, which is cast from a 3D-printed mould. Droplets of 0.7-6.0 μL volume were packed as homogeneous or heterogeneous linear arrays of 2-9 droplets that were stable for at least six hours. The interdroplet bilayers had an area of up to 0.56 mm 2 , or an equivalent diameter of up to 850 μm, as determined from capacitance measurements. We observed osmotic water transfer over the bilayers as well as sequential bilayer lysis by the pore-forming toxin melittin. These millifluidic interdroplet bilayer arrays combine the ease of electrical and optical access of manually pipetted microdroplets with the automation and reproducibility of microfluidic technologies. Moreover, the 3D-printing based fabrication strategy enables the rapid implementation of alternative channel geometries, e.g. branched arrays, with a design-to-device time of just 24-48 hours.

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“…The peptide spontaneously forms an α-helix upon insertion in a lipid membrane and is well known to induce leakage from liposomes [15]. The ability of disk-associated melittin to detach and redistribute into another lipid membrane was investigated by monitoring liposome leakage after addition of peptide-lipid disk mixtures.…”

Section: Liposome Leakagementioning

confidence: 99%

“…These findings, together with the biocompatible and stable properties of the disks, lead us to propose PEG-stabilized lipid disks as potential carriers for amphiphilic antimicrobial peptides. In this study we use melittin [15] as a model peptide to further investigate the possibility of using the disks for formulation of antimicrobial peptides. One question we set out to answer is whether the peptides bound to the lipid disk are able to detach from the carrier and redistribute into a target membrane.…”

Section: Introductionmentioning

confidence: 99%

PEG-stabilized lipid disks as carriers for amphiphilic antimicrobial peptides

Zetterberg

Reijmar

Pränting

et al. 2011

Journal of Controlled Release

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This is an accepted version of a paper published in Journal of Controlled Release. This paper has been peer-reviewed but does not include the final publisher proof-corrections or journal pagination.Citation for the published paper: Zetterberg, M., Reijmar, K., Pränting, M., Engström, Å., Andersson, D. et al. (2011) AbstractAntimicrobial peptides hold potential as a possible alternative, or complement, to conventional antibiotics but new, safe and efficient means are needed for formulation and administration of the peptides. In this study we have investigated the utility of a novel type of lipid particles, the polyethylene glycol-stabilized lipid disks, as carriers for the model peptide melittin. The structural integrity of the carrier particle when loaded with the peptide was investigated using cryo-transmission electron microscopy. Liposome leakage upon addition of the peptide-lipid disks was monitored as a means to verify the membrane lytic effect of the formulation. The susceptibility of melittin to tryptic digestion was studied and compared in the absence and presence of lipid disks. Finally, the antibacterial effect of the peptide-lipid disk formulation was compared to that of free melittin after both single and repeated exposure to Escherichia coli. The results show that melittin can redistribute from the disk into a new host membrane and that formulation in the disks does not compromise melittin's membrane permeabilizing ability. Further, the peptide was found to be fully protected against degradation when bound to the disks. Time-kill experiments revealed that all the antibacterial effect of melittin administered in free form was gone after a single exposure to E. coli. In contrast, the disk formulation showed significant cell-killing effect also upon a second exposure to bacteria, indicating an extended release of peptide from the lipid disks. These results suggest that the lipid disks constitute a new class of promising carriers for peptide antibiotics.

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Melittin: a Membrane-active Peptide with Diverse Functions

Cited by 539 publications

References 267 publications

High content analysis to determine cytotoxicity of the antimicrobial peptide, melittin and selected structural analogs

High content analysis to determine cytotoxicity of the antimicrobial peptide, melittin and selected structural analogs

Interdroplet bilayer arrays in millifluidic droplet traps from 3D-printed moulds

PEG-stabilized lipid disks as carriers for amphiphilic antimicrobial peptides

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