Structural Characterization of the Antimicrobial Peptide Pleurocidin from Winter Flounder

Syvitski, Raymond T.; Burton, Ian W.; Mattatall, Neil R.; Douglas, Susan E.; Jakeman, David L.

doi:10.1021/bi0504005

Cited by 64 publications

(73 citation statements)

References 77 publications

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“…It is also desirable that such a fusion peptide should be relatively stable and retain its bactericidal activity under a range of physiological conditions, such as different salt concentrations, ionic strengths, pHs, and other factors in body fluids (8,15,34). By studying marine-derived pleurocidin variants (25,30) and an S. mutans quorum-sensing signaling peptide pheromone or CSP (17, 18, 31) we have The peptide bands were quantitated by densitometry to calculate percent degradation compared to the 0-min samples. Lane 1, low-molecular-weight markers; lane 2, IMB-2 only (ϳ4.5 kDa) as a blank control; lanes 3 to 6, IMB-2 exposed to saliva with 1 mM EDTA for 0, 5, 10, and 15 min; lanes 7 to 10, IMB-2 exposed to saliva without EDTA for 0, 5, 10, and 15 min.…”

Section: Discussionmentioning

confidence: 99%

A Novel Target-Specific, Salt-Resistant Antimicrobial Peptide against the Cariogenic Pathogen Streptococcus mutans

Mai

Tian

Gallant

et al. 2011

Antimicrob Agents Chemother

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In this study, we constructed and evaluated a target-specific, salt-resistant antimicrobial peptide (AMP) that selectively targeted Streptococcus mutans, a leading cariogenic pathogen. The rationale for creating such a peptide was based on the addition of a targeting domain of S. mutans ComC signaling peptide pheromone (CSP) to a killing domain consisting of a portion of the marine-derived, broad-spectrum AMP pleurocidin to generate a target-specific AMP. Here, we report the results of our assessment of such fusion peptides against S. mutans and two closely related species. The results showed that nearly 95% of S. mutans cells lost viability following exposure to fusion peptide IMB-2 (5.65 M) for 15 min. In contrast, only 20% of S. sanguinis or S. gordonii cells were killed following the same exposure. Similar results were also observed in dual-species mixed cultures of S. mutans with S. sanguinis or S. gordonii. The peptide-guided killing was further confirmed in S. mutans biofilms and was shown to be dose dependent. An S. mutans mutant defective in the CSP receptor retained 60% survival following exposure to IMB-2, suggesting that the targeted peptide predominantly bound to the CSP receptor to mediate killing in the wild-type strain. Our work confirmed that IMB-2 retained its activity in the presence of physiological or higher salt concentrations. In particular, the fusion peptide showed a synergistic killing effect on S. mutans with a preventive dose of NaF. In addition, IMB-2 was relatively stable in the presence of saliva containing 1 mM EDTA and did not cause any hemolysis. We also found that replacement of serine-14 by histidine improved its activity at lower pH. Because of its effectiveness, salt resistance, and minimal toxicity to host cells, this novel target-specific peptide shows promise for future development as an anticaries agent.

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Section: Discussionmentioning

confidence: 99%

A Novel Target-Specific, Salt-Resistant Antimicrobial Peptide against the Cariogenic Pathogen Streptococcus mutans

Mai

Tian

Gallant

et al. 2011

Antimicrob Agents Chemother

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“…The last few residues at the C termini have slightly more motional freedom than the core of the helix. We dissolved the peptides in the widely used solvent TFE, because it improves the structural stability of peptides and is considered to be biologically relevant (7,41). TFE is also thought to mimic a protein receptor environment by stabilizing helices in regions with intrinsic ␣-helical propensity that are likely to form helices when they bind to their protein partner (36,44).…”

Section: Discussionmentioning

confidence: 99%

“…S3 and S4 in the supplemental material). Sequence-specific assignments were determined by the methods described previously (41). Only i to iϩ1, i to iϩ3, and i to iϩ4 interresidue nuclear Overhauser effect spectroscopy (NOESY) cross-peaks were observed for residues between Leu4 and Gly20 of UA159sp and between Ser5 and Thr16 of TPC3 (see Fig.…”

Section: Methodsmentioning

confidence: 99%

“…Two series of structural calculations were performed with and without added hydrogen bonds to assess the effect of adding suspected hydrogen bonds on the structure calculation. All structural calculations were based on previous studies (10,34,41), using the XPLOR 3.1 software package. A total of 33 and 21 UA159sp lowest-energy structures (with and without hydrogen bonds included in the simulation) and 43 and 36 TPC3 lowest-energy structures (with and without hydrogen bonds included in the simulation) were retained, which had no violations of NOESY constraints of Ͼ0.5 Å .…”

Section: Methodsmentioning

confidence: 99%

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Structure-Activity Analysis of Quorum-Sensing Signaling Peptides from Streptococcus mutans

Syvitski¹,

Tian²,

Sampara³

et al. 2007

J Bacteriol

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Streptococcus mutans secretes and utilizes a 21-amino-acid signaling peptide pheromone to initiate quorum sensing for genetic competence, biofilm formation, stress responses, and bacteriocin production. In this study, we designed and synthesized a series of truncated peptides and peptides with amino acid substitutions to investigate their structure-activity relationships based on the three-dimensional structures of S. mutans wild-type signaling peptide UA159sp and C-terminally truncated peptide TPC3 from mutant JH1005 defective in genetic competence. By analyzing these peptides, we demonstrated that the signaling peptide of S. mutans has at least two functional domains. The C-terminal structural motif consisting of a sequence of polar hydrophobic charged residues is crucial for activation of the signal transduction pathway, while the core ␣-helical structure extending from residue 5 to the end of the peptide is required for receptor binding. Peptides in which three or more residues were deleted from the C terminus did not induce genetic competence but competitively inhibited quorum sensing activated by UA159sp. Disruption of the amphipathic ␣-helix by replacing the Phe-7, Phe-11, or Phe-15 residue with a hydrophilic residue resulted in a significant reduction in or complete loss of the activity of the peptide. In contrast to the C-terminally truncated peptides, these peptides with amino acid substitutions did not compete with UA159sp to activate quorum sensing, suggesting that disruption of the hydrophobic face of the ␣-helical structure results in a peptide that is not able to bind to the receptor. This study is the first study to recognize the importance of the signaling peptide C-terminal residues in streptococcal quorum sensing.

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“…For all samples, the concentration of peptide was 5 mM. One-and two-dimensional 1 H NMR data sets were collected on a Bruker AVANCE 500 spectrometer and processed as previously described (33). For structure calculations, distance restraints determined from the integration of NOESY cross-peaks (80 ms for myristoylated p15 peptide and 250 ms for non-myristoylated p15 peptide) were classified into four groups: strong, medium, weak, and very weak, corresponding to inter-proton distance ranges of Ͻ2.3, 2.0 -3.5, 3.3-5.0, and 4.8 -6.0 Å, respectively.…”

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confidence: 99%

Cell-Cell Membrane Fusion Induced by p15 Fusion-associated Small Transmembrane (FAST) Protein Requires a Novel Fusion Peptide Motif Containing a Myristoylated Polyproline Type II Helix

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Dawe

et al. 2012

Journal of Biological Chemistry

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Background:The p15 FAST protein mediates cell-cell fusion using a 19-residue ectodomain. Results: The p15 ectodomain requires both an N-terminal myristate and a polyproline type II helix for membrane fusion activity. Conclusion:The p15 ectodomain functions as a novel fusion peptide motif. Significance: This novel fusion peptide provides new insights into the essential biological process of protein-mediated membrane fusion.

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Structural Characterization of the Antimicrobial Peptide Pleurocidin from Winter Flounder

Cited by 64 publications

References 77 publications

A Novel Target-Specific, Salt-Resistant Antimicrobial Peptide against the Cariogenic Pathogen Streptococcus mutans

A Novel Target-Specific, Salt-Resistant Antimicrobial Peptide against the Cariogenic Pathogen Streptococcus mutans

Structure-Activity Analysis of Quorum-Sensing Signaling Peptides from Streptococcus mutans

Cell-Cell Membrane Fusion Induced by p15 Fusion-associated Small Transmembrane (FAST) Protein Requires a Novel Fusion Peptide Motif Containing a Myristoylated Polyproline Type II Helix

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