Analysis of antimicrobial peptide interactions with hybrid bilayer membrane systems using surface plasmon resonance

Mozsolits, Henriette; Wirth, Hans-Jürgen; Werkmeister, Jerome A.; Aguilar, Marie Isabel

doi:10.1016/s0005-2736(01)00303-0

Cited by 160 publications

(203 citation statements)

References 45 publications

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“…The present results therefore reveal that the C-terminal charges of melittin reduce the affinity of the peptide for neutral membranes. These conclusions are in contradiction with those inferred from the SPR binding studies concluding that truncated amidated melittin-21Q exhibits, relative to melittin, a lower or a similar affinity for DMPC membranes 45,46 . These SPR experiments were conducted on supported bilayers and the nature …”

Section: Acs Paragon Plus Environmentcontrasting

confidence: 92%

“…For instance, the properties of melittin-21Q, a truncated analogue of melittin in which residues 21 to 25 (K 21 -R 22 -K 23 -R 24 -Q 25 ) were omitted, were investigated. Its binding to zwitterionic 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) bilayers as well as to anionic 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG) bilayers was measured by Surface Plasmon Resonance (SPR) [45][46] . Melittin-21Q exhibited a reduced or similar binding to DMPC and to DMPG bilayers compared to native melittin, suggesting a favorable or a limited contribution of the cationic C-terminal portion to membrane affinity.…”

Section: Introductionmentioning

confidence: 99%

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Role of the Cationic C-Terminal Segment of Melittin on Membrane Fragmentation

2016

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The widespread distribution of cationic antimicrobial peptides capable of membrane fragmentation in nature underlines their importance to living organisms. In the present work, we determined the impact of the electrostatic interactions associated with the cationic Cterminal segment of melittin, a 26-amino acid peptide from bee venom (net charge +6), on its binding to model membranes and on the resulting fragmentation. In order to detail the role played by the C-terminal charges, we prepared a melittin analogue for which the 4 cationic amino acids in positions 21 to 24 were substituted with the polar residue citrulline, providing a peptide with the same length and amphiphilicity but with a lower net charge (+2). We compared the peptide bilayer affinity and the membrane fragmentation for bilayers prepared from dipalmitoyl-sn-glycero-3-phosphocholine (DPPC)/1,2-dipalmitoyl-sn-glycero-3-phospho-L-serine (DPPS) mixtures. It is shown that neutralization of the C-terminal considerably increased melittin affinity for zwitterionic membranes. The unfavorable contribution associated with transferring the cationic C-terminal in a less polar environment was reduced, leaving the hydrophobic interactions, which drive the peptide insertion in bilayers, with limited counterbalancing interactions. The presence of negatively charged lipids (DPPS) in bilayers increased melittin binding by introducing attractive electrostatic interactions, the augmentation being, as expected, greater for native melittin than for its citrullinated analogue. The membrane fragmentation power of the peptide was shown to be controlled by electrostatic interactions and could be modulated by the charge carried by both the membrane and the lytic peptide. The analysis of the lipid composition of the extracted fragments from DPPC/DPPS bilayers revealed no lipid specificity. It is proposed that extended phase separations are more susceptible to lead to the extraction of a lipid species in a specific manner than a specific lipid-peptide affinity. The present work on the lipid extraction by melittin and citrullinated melittin with model membranes emphasizes the complex relation between the affinity, the lipid extraction/membrane fragmentation, and the lipid specificity.

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Section: Acs Paragon Plus Environmentcontrasting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Role of the Cationic C-Terminal Segment of Melittin on Membrane Fragmentation

2016

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“…We have already established an evaluation method of the interactions of various peptides to lipid membranes using SPR, [9][10][11][12][13][14][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.…”

Section: Introductionmentioning

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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“…The corresponding differential rate equations for this reaction model are represented by [18][19][20][21][22][23] …”

Section: Discussionmentioning

confidence: 99%