Interaction of the Antimicrobial Peptide Gomesin with Model Membranes: A Calorimetric Study

Domingues, Tatiana M.; Mattei, Bruno; Seelig, Joachim; Perez, Kátia Regina; Miranda, Antônio; Riske, Karin A.

doi:10.1021/la401596s

Cited by 45 publications

(101 citation statements)

References 55 publications

(100 reference statements)

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“…ITC data showed that binding of Gm to anionic membranes is an exothermic process that follows a stoichiometry of roughly one gomesin charge per lipid charge. Light scattering data show that Gm binding is always accompanied by peptide-induced lipid aggregation [35]. This phenomenon is not exclusive for Gm, and it was reported for other antimicrobial peptides [37,38].…”

Section: Accepted Manuscriptmentioning

confidence: 55%

“…The activity of GmL was found to be significantly lower than that of Gm for low POPG fraction, but becomes comparable to that of Gm for high POPG content. In recent works, we studied the interaction of Gm and different analogues that preserve the -hairpin conformation with large unilamellar vesicles (LUVs) composed of POPC and POPG using different approaches, namely isothermal titration calorimetry (ITC), leakage of a fluorescent probe entrapped in vesicles and light scattering measurements [35,36]. ITC data showed that binding of Gm to anionic membranes is an exothermic process that follows a stoichiometry of roughly one gomesin charge per lipid charge.…”

Section: Accepted Manuscriptmentioning

confidence: 99%

“…The CF leakage percentage was calculated using the following equation: %Leakage = 100(F t -F 0 )/(F max -F 0 ), where F 0 is the CF fluorescence before peptide addition, F t is the CF fluorescence at the chosen time, and F max is the CF fluorescence after detergent addition [35].…”

Section: Preparation Of Large Unilamellar Vesicles (Luvs)mentioning

confidence: 99%

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Comparative study of the mechanism of action of the antimicrobial peptide gomesin and its linear analogue: The role of the β-hairpin structure

Domingues

Perez

Miranda

et al. 2015

Biochimica et Biophysica Acta (BBA) - Biomembranes

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Gomesin (Gm) is an antimicrobial peptide first isolated from the hemolymph of a Brazilian spider. Its powerful antimicrobial activity is, however, accompanied by hemolysis. As an alternative to this issue, a linear analogue (named GmL) lacking the disulfide bonds was designed. Here, CD spectroscopy, a fluorescence-based leakage assay, isothermal titration calorimetry (ITC) and light scattering are used to study the interaction of both Gm and GmL with large unilamellar vesicles (LUVs) composed of POPC (palmitoyl oleoyl phosphatidylcholine) with 25 and 50 mol% POPG (palmitoyl oleoyl phosphatidylglycerol). The activities of Gm and GmL in respect to their binding affinity/enthalpy, ability to permeabilize membranes and to induce vesicle aggregation are correlated with peptide secondary structure. Whereas Gm displays a quite stable β-hairpin motif irrespective of the environment, GmL assumes a random conformation in aqueous solution and in the presence of 25 mol% POPG but adopts a β-like structure in the presence of 50 mol% POPG. Gm exhibited high lytic activity against both surface charge densities. Instead, the activity of GmL was found to be negligible in the presence of 25 mol% POPG LUVs, but comparable to that of the native peptide against 50 mol% POPG as a consequence of peptide structuring. We conclude that the activity of Gm and its linear analogue is intimately related to the formation of a β-turn motif, in which the hydrophobic residues form a hydrophobic face able to insert into the membrane and disrupt it.

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Section: Accepted Manuscriptmentioning

confidence: 55%

Section: Accepted Manuscriptmentioning

confidence: 99%

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Comparative study of the mechanism of action of the antimicrobial peptide gomesin and its linear analogue: The role of the β-hairpin structure

Domingues

Perez

Miranda

et al. 2015

Biochimica et Biophysica Acta (BBA) - Biomembranes

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“…Using the assumption that binding heat is constant, the number of bound ligands can be determined from the total heat change. ITC allows the calculation of the binding constant and enthalpic and entropic changes of the binding process [199,205]. An important advantage of ITC is that it can be performed with unmodified, native forms of molecules, hence it does not introduce artifacts [206,207].…”

mentioning

confidence: 99%

“…Furthermore, ITC is a preferred approach for the analysis of CPPs that are coupled to larger biomolecules since CD and NMR spectra of these large molecules are very difficult to analyze [208] and unlike NMR techniques, ITC does not require large amounts of samples [209]. There are different applications of ITC to characterize the thermodynamic aspects of peptide-membrane interactions [205,[210][211][212][213][214].…”

mentioning

confidence: 99%

Membrane Active Peptides and Their Biophysical Characterization

Avcı

Akbulut

Özkırımlı

2018

Preprint

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In the last 20 years, an increasing number of studies have been reported on membrane active peptides, which exert their biological activity by interacting with the cell membrane either to disrupt it and lead to cell lysis or to translocate through it to deliver cargos into the cell and reach their target. These peptides are attractive alternatives to currently used pharmaceuticals. Antimicrobial peptides (AMPs) and peptides designed for drug and gene delivery currently in the drug pipeline suggest that these membrane active peptides will soon constitute a significant percentage of the drug market. Here, we focus on two most prominent classes of membrane active peptides; AMPs and cell-penetrating peptides (CPPs). AMPs are a group of membrane active peptides that disrupt the membrane integrity or inhibit the cellular functions of bacteria, virus and fungi. CPPs are another group of membrane active peptides that mainly function as cargo-carriers even though they may also show antimicrobial activity to some extent. Biophysical techniques to understand how they interact with the membrane have shed light on the peptide-membrane interaction at various levels of detail. Structural investigation of membrane active peptides in the presence of the membrane provides important clues on the effect of the membrane environment on peptide conformations. Advances in live imaging techniques have allowed examination of peptide action at a single cell or single molecule level. In addition to these experimental biophysical techniques, molecular dynamics simulations provided clues on the peptide-lipid interactions and dynamics of the cell entry process at atomic detail. In this review, we summarize the recent advances in experimental and computational investigation of membrane active peptides with particular emphasis on two amphipathic membrane active peptides, the AMP melittin and the CPP pVEC.

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Structure‐activity relationship of Trp‐containing analogs of the antimicrobial peptide gomesin

Domingues

Buri

Daffre

et al. 2014

Journal of Peptide Science

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Gomesin (Gm) has a broad antimicrobial activity making it of great interest for development of drugs. In this study, we analyzed three Gm analogs, [Trp(1) ]-Gm, [Trp(7) ]-Gm, and [Trp(9) ]-Gm, in an attempt to gain insight into the contributions of different regions of the peptide sequence to its activity. The incorporation of the tryptophan residue in different positions has no effect on the antimicrobial and hemolytic activities of the Gm analogs in relation to Gm. Spectroscopic studies (circular dichroism, fluorescence and absorbance) of Gm and its analogs were performed in the presence of SDS, below and above its critical micelle concentration (CMC) (~8 mM), in order to monitor structural changes induced by the interaction with this anionic surfactant (0-15 mM). Interestingly, we found that the analogs interact more strongly with SDS at low concentrations (0.3-6.0 mM) than close to or above its CMC. This suggests that SDS monomers are able to cover the whole peptide, forming large detergent-peptide aggregates. On the other hand, the peptides interact differently with SDS micelles, inserting partially into the micelle core. Among the peptides, Trp in position 1 becomes more motionally-restricted in the presence of SDS, probably because this residue is located at the N-terminal region, which presents higher conformational freedom to interact stronger with SDS molecules. Trp residues in positions 7 and 9, close to and in the region of the turn of the molecule, respectively, induced a more constrained structure and the compounds cannot insert deeper into the micelle core or be completely buried by SDS monomers.

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Interaction of the Antimicrobial Peptide Gomesin with Model Membranes: A Calorimetric Study

Cited by 45 publications

References 55 publications

Comparative study of the mechanism of action of the antimicrobial peptide gomesin and its linear analogue: The role of the β-hairpin structure

Comparative study of the mechanism of action of the antimicrobial peptide gomesin and its linear analogue: The role of the β-hairpin structure

Membrane Active Peptides and Their Biophysical Characterization

Structure‐activity relationship of Trp‐containing analogs of the antimicrobial peptide gomesin

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