Membrane Association, Electrostatic Sequestration, and Cytotoxicity of Gly-Leu-Rich Peptide Orthologs with Differing Functions

Vanhoye, Damien; Bruston, Francine; Amri, Shaharazade El; Ladram, Ali; Amiche, Mohamed; Nicolas, Pierre

doi:10.1021/bi0493158

Cited by 49 publications

(69 citation statements)

References 83 publications

(121 reference statements)

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“…Resins and Fmoc-protected amino acids were purchased from Senn Chemicals (Switzerland) and solvents from sds (France) as previously described. 1 Briefly, synthesis products were cleaved from the resin (Wang PS resin) by a mixture of trifluoracetic acid (TFA) (95%), H 2 O (2.5%), and Triisopropylsilan (2.5%), precipitated in ether, centrifuged, then lyophilized. The peptides were purified by a semi-preparative HPLC (C18 For sake of simplicity of mention DRP will be omitted when peptides are cited in the text.…”

Section: Synthesis and Purificationmentioning

confidence: 99%

“…They have very similar amino acid sequences, hydrophobicities, and amphipathicities (Table I), but differ markedly in their membrane-damaging properties. These include the cationic peptide DRP-PBN2, 1 GLVTSLIKGAGKLLGGLFGSVTG-NH 2 , designated PBN2KF, as it contains two lysines (positions 8 and 12) and one phe-…”

Section: Introductionmentioning

confidence: 99%

“…Although PBN2KF is hemolytic and very potent against microorganisms, PD36 has only hemolytic activity. 1 Previously, we have used ancestral protein reconstruction and peptide analogue design to examine the roles of electrostatic and hydrophobic interactions in the biological activity and mode of action of these functionally divergent GL-rich peptides. PD36K and PD36KF analogues of PD36 were synthesized by replacing residues at positions 8 and 12 with K, and at position 18 with F 1,3 ( Table I).…”

Section: Introductionmentioning

confidence: 99%

“…The data demonstrated that evolution has positively selected charge-altering nucleotide substitutions to generate from an otherwise neutral ancestral hemolytic peptide orthologous cationic variants that bind to and permeate bacterial cell membranes. 1,3 This process leads to the selection of lysine residues. In a recent work, we have underlined the impact of structural polymorphism of plasticins on adsorption process onto mimetic membranes, lipid packing disturbance, and antimicrobial or cytotoxic activities.…”

Section: Introductionmentioning

confidence: 99%

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Structural malleability of plasticins: Preorganized conformations in solution and relevance for antimicrobial activity

et al. 2007

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Plasticins (23 long-residue glycine-leucine-rich dermaseptin-related peptides produced by the skin of South American hylids) have very similar amino acid sequences, hydrophobicities, and amphipathicities, but differ in their membrane-damaging properties and structurations (i.e. destabilized helix states, beta-hairpin, beta-sheet, and disordered states) at anionic and zwitterionic membrane interfaces. Structural malleability of plasticins in aqueous solutions together with parameters that may govern their ability to fold within beta-hairpin like structures were analyzed through circular dichroism and FTIR spectroscopic studies completed by molecular dynamics simulations in polar mimetic media. The goal of this study was to probe to which extent pre-existent peptide conformations, i.e. intrinsic "conformational landscape", may be responsible for variability in bioactive conformation and antimicrobial/hemolytic mechanisms of action of these peptides in relation with their various membrane disturbing properties. All plasticins present a turn region that does not always result in folding into a beta-hairpin shaped conformation. Residue at position 8 plays a major role in initiating the folding, while position 12 is not critical. Conformational stability has no major impact on antimicrobial efficacy. However, preformed beta-hairpin in solution may act as a conformational lock that prevents switch to alpha-helical structure. This lock lowers the antimicrobial efficiency and explains subtle differences in potencies of the most active antimicrobial plasticins.

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Section: Synthesis and Purificationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Structural malleability of plasticins: Preorganized conformations in solution and relevance for antimicrobial activity

et al. 2007

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“…This model was chosen based on previous studies of peptides-membrane interaction. [9][10][11][12][13][14][15][24][25][26][27] …”

Section: Peptide Binding To the Bilayer Membranementioning

confidence: 99%

Evaluation of the Membrane-binding Properties of the Proximal Region of the Angiotensin II Receptor (AT1A) Carboxyl Terminus by Surface Plasmon Resonance

et al. 2005

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IntroductionG protein-coupled receptors (GPCR) play an important role in signaling cascades, where they participate in the transduction of extracellular signals to intracellular heterotrimeric guanyl nucleotide binding proteins (G proteins). GPCRs are a large superfamily with a structure comprising an extracellular amino terminus, seven transmembrane-spanning α-helices connected by alternating extracellular and intracellular loops, and a cytoplasmic carboxyl terminal region.1 The arrangement of the seven transmembrane-spanning helices and the extracellular domains provides a specific binding site for ligands, which induces a conformational change in the receptor that exposes intracellular regions, which recruit and activate G proteins.The type 1 angiotensin II (AngII) receptor (AT1A) is a 359 amino acid GCPR that mediates the important cardiovascular and homeostatic actions of the peptide hormone angiotensin II. The 54 amino acid intracellular carboxyl terminus (Leu 305 to Glu 359 ) of the AT1A receptor interacts with G proteins 2 and other signaling molecules, 3 indicating a contribution to receptor activation, while the discovery of phosphorylation sites 4 and internalization motifs 5-7 suggest an involvement in receptor regulation. In particular, the proximal region (Leu 305 to Pro 321 ), comprising the first third of the AT1A receptor carboxylterminus, is an important site of complex interactions for receptor function.In previous work, we proposed that the proximal carboxylterminus of the AT1A receptor (residues 305 -320) is an amphipathic α-helix (now referred to as helix VIII), and that this structure may be relevant to receptor activation and internalization on the basis of computer modeling. 6,8,9 In order to investigate the conformation and orientation of the proximal carboxyl terminus, the interaction of a synthetic peptide corresponding to the carboxyl terminus (Leu 305 to Lys 325 ) of the receptor (AT peptide) and its analogue peptide (where lysine residues at 307, 308, 310, and 311 were substituted with norleucine) with a zwitterionic or anionic lipid membrane as a model was examined by surface plasmon resonance (SPR). 8 The AT peptide was shown by SPR to bind with high affinity to a negatively charged lipid membrane via electrostatic interactions.We have already established an evaluation method of the interactions of various peptides to lipid membranes using SPR, 9-15 and demonstrated that the kinetic rate constants and the affinity of membrane interactions can be readily determined by this technique, which in turn provides important insights into the mechanism of peptide-membrane interactions. In the present study, we extended our studies into the membrane binding properties of the AT peptide.Specifically, we examined the membrane-binding properties of a series of peptide analogues of the AT peptide in which particular amino acid residues have been modified to establish their role in the structure and membrane binding. We determined the kinetic rate constants and affinity of these synthetic peptid...

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Effect of membrane mimicking environment on the conformation of a pore‐forming (xSxG)₆peptide

2006

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The mechanism of membrane interaction by beta-sheet peptides is important to understand fundamental principles of folding of beta-barrel proteins and various beta-amyloid proteins. Here, we examined the conformational characteristics of a porin-like channel forming (xSxG)(6) peptide in solution and membrane-mimicking environments (CD and ATR-IR) to understand the structural changes of the peptide during membrane association and channel formation. A comparison of the peptide conformations in different microenvironments showed that beta-sheet formation is enhanced in membrane-mimicking liposomes and SDS-micelles. The lipid-induced beta-sheet formation was confirmed by the formation of a characteristic beta-sheet structure on mixing a methanolic solution of the peptide (partially folded) with preformed liposomes. The amphipathicity of the peptide; increased hydrogen bonding, hydrophilicity, and reduction in dimensionality of the membrane surface; membrane-peptide interaction-forces; and presence of flexible glycines might facilitate beta-sheet formation in membranes. Though the CD spectra of both the peptide-bound and peptide-incorporated lipids are reminiscent of a beta-sheet structure, a significant variation in the peak positions of the two beta-sheet structures was noticed. The channel characteristics of (xSxG)(6) in the presence of low ionic strength solutions of NEt(3)BzCl and glucosammonium chloride are comparable to those reported under high ionic strength solutions. Altogether the data suggest that the channel formation by (xSxG)(6) proceeds via beta-sheet aggregate formation at the membrane surface, beta-sheet insertion, and rearrangement into a beta-barrel-like structure. The beta-barrel-like channel formation most likely arises from a sequence similarity to beta-barrel porins whereas the lipid-induced beta-sheet formation is governed by the above-mentioned factors.

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Membrane Association, Electrostatic Sequestration, and Cytotoxicity of Gly-Leu-Rich Peptide Orthologs with Differing Functions

Cited by 49 publications

References 83 publications

Structural malleability of plasticins: Preorganized conformations in solution and relevance for antimicrobial activity

Structural malleability of plasticins: Preorganized conformations in solution and relevance for antimicrobial activity

Evaluation of the Membrane-binding Properties of the Proximal Region of the Angiotensin II Receptor (AT1A) Carboxyl Terminus by Surface Plasmon Resonance

Effect of membrane mimicking environment on the conformation of a pore‐forming (xSxG)₆peptide

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Membrane Association, Electrostatic Sequestration, and Cytotoxicity of Gly-Leu-Rich Peptide Orthologs with Differing Functions

Cited by 49 publications

References 83 publications

Structural malleability of plasticins: Preorganized conformations in solution and relevance for antimicrobial activity

Structural malleability of plasticins: Preorganized conformations in solution and relevance for antimicrobial activity

Evaluation of the Membrane-binding Properties of the Proximal Region of the Angiotensin II Receptor (AT1A) Carboxyl Terminus by Surface Plasmon Resonance

Effect of membrane mimicking environment on the conformation of a pore‐forming (xSxG)6peptide

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Effect of membrane mimicking environment on the conformation of a pore‐forming (xSxG)₆peptide