Journal: ACS Omega

Article Title: Cell-Selective Pore Forming Antimicrobial Peptides of the Prodomain of Human Furin: A Conserved Aromatic/Cationic Sequence Mapping, Membrane Disruption, and Atomic-Resolution Structure and Dynamics

doi: 10.1021/acsomega.8b01876

Figure Lengend Snippet: Furin prodomain-derived antibacterial peptides in human RBC lysis, mouse fibroblast cell NIH3T3 viability, and cytokine release from human THP-1 cell line. (A) Estimated % lysis of RBC as a function of concentrations of YR26, YR23, YR20, YR12, and RV23 peptides. (B) Peptides dose-dependent viability of mouse fibroblast cells. (C, D) Dose-dependent inhibition of (C) TNF-α and (D) IL-1β secretion by the YR23 peptide from LPS-induced THP-1 cells.

Article Snippet: A synthetic gene of YR26, from Shanghai ShineGene Molecular Biotech, Inc, was cloned as a fusion protein in a pET14b vector containing hydrophobic KSI at the N-terminus.

Techniques: Derivative Assay, Red Blood Cell Lysis, Lysis, Inhibition

Journal: ACS Omega

Article Title: Cell-Selective Pore Forming Antimicrobial Peptides of the Prodomain of Human Furin: A Conserved Aromatic/Cationic Sequence Mapping, Membrane Disruption, and Atomic-Resolution Structure and Dynamics

doi: 10.1021/acsomega.8b01876

Figure Lengend Snippet: Prodomain-derived antibacterial peptides disrupted the outer membrane and neutralized the surface charge of E. coli cells. (A) Outer membrane permeability assay using NPN as a fluorescence probe. Increase in fluorescence intensity by the antibacterial peptides, YR26, YR23, and YR20 indicated their efficacy to permeabilize the outer cell membrane. (B) The ζ-potential of E. coli cells in the presence of varying concentrations of furin prodomain-derived peptides.

Article Snippet: A synthetic gene of YR26, from Shanghai ShineGene Molecular Biotech, Inc, was cloned as a fusion protein in a pET14b vector containing hydrophobic KSI at the N-terminus.

Techniques: Derivative Assay, Membrane, Permeability, Fluorescence

Journal: ACS Omega

Article Title: Cell-Selective Pore Forming Antimicrobial Peptides of the Prodomain of Human Furin: A Conserved Aromatic/Cationic Sequence Mapping, Membrane Disruption, and Atomic-Resolution Structure and Dynamics

doi: 10.1021/acsomega.8b01876

Figure Lengend Snippet: Binding energetics of prodomain-derived peptides in complex LPS. ITC thermograms of peptides (A) YR26, (B) YR23, (C) YR20, and (D) YR12. The top panel of each ITC profile shows titrated peaks as a function of time and the lower panel represents the integrated heat value of each titration point. ITC experiments were carried out in sodium phosphate buffer, pH 7.0, 37 °C.

Article Snippet: A synthetic gene of YR26, from Shanghai ShineGene Molecular Biotech, Inc, was cloned as a fusion protein in a pET14b vector containing hydrophobic KSI at the N-terminus.

Techniques: Binding Assay, Derivative Assay, Titration

Journal: ACS Omega

Article Title: Cell-Selective Pore Forming Antimicrobial Peptides of the Prodomain of Human Furin: A Conserved Aromatic/Cationic Sequence Mapping, Membrane Disruption, and Atomic-Resolution Structure and Dynamics

doi: 10.1021/acsomega.8b01876

Figure Lengend Snippet: Interactions and perturbation of LPS-induced by prodomain-derived antibacterial peptides. Intrinsic Trp fluorescence experiments of YR26, YR23, YR20, and YR12 peptides as a function of concentrations of LPS. (A) Changes in fluorescence intensity at the emission maxima and (B) changes in emission wavelength (λ max ). (C–F) Bar diagrams show particle size distribution of LPS, estimated from DLS experiments, in free solutions and at different molar ratios of peptides (C) YR26, (D) YR23, (E) YR20, and (F) YR12.

Article Snippet: A synthetic gene of YR26, from Shanghai ShineGene Molecular Biotech, Inc, was cloned as a fusion protein in a pET14b vector containing hydrophobic KSI at the N-terminus.

Techniques: Derivative Assay, Fluorescence

Journal: ACS Omega

Article Title: Cell-Selective Pore Forming Antimicrobial Peptides of the Prodomain of Human Furin: A Conserved Aromatic/Cationic Sequence Mapping, Membrane Disruption, and Atomic-Resolution Structure and Dynamics

doi: 10.1021/acsomega.8b01876

Figure Lengend Snippet: Binding of prodomain-derived antibacterial peptides to lipid vesicles. Estimation of CF dye release as a function of concentrations of YR26, YR23, and YR12 peptides from (A) POPC–POPG vesicles and (B) POPC vesicles. (C–F) Bar diagrams show the particle size distribution of POPC–POPG vesicles, estimated from DLS experiments, in free solutions and at different molar ratios of peptides (C) YR26, (D) YR23, (E) YR20, and (F) YR12.

Article Snippet: A synthetic gene of YR26, from Shanghai ShineGene Molecular Biotech, Inc, was cloned as a fusion protein in a pET14b vector containing hydrophobic KSI at the N-terminus.

Techniques: Binding Assay, Derivative Assay

Journal: ACS Omega

Article Title: Cell-Selective Pore Forming Antimicrobial Peptides of the Prodomain of Human Furin: A Conserved Aromatic/Cationic Sequence Mapping, Membrane Disruption, and Atomic-Resolution Structure and Dynamics

doi: 10.1021/acsomega.8b01876

Figure Lengend Snippet: NMR analyses of the antibacterial YR26 peptide fragment of prodomain of furin in SDS micelles. (A) The 15 N– 1 H HSQC spectrum of 15 N-labeled YR26 peptide showing assignment of individual residue. (B) Bar diagram delineates the number of NOE contacts for each residue of YR26 peptide.

Article Snippet: A synthetic gene of YR26, from Shanghai ShineGene Molecular Biotech, Inc, was cloned as a fusion protein in a pET14b vector containing hydrophobic KSI at the N-terminus.

Techniques: Labeling, Residue

Journal: ACS Omega

Article Title: Cell-Selective Pore Forming Antimicrobial Peptides of the Prodomain of Human Furin: A Conserved Aromatic/Cationic Sequence Mapping, Membrane Disruption, and Atomic-Resolution Structure and Dynamics

doi: 10.1021/acsomega.8b01876

Figure Lengend Snippet: Atomic-resolution structure of the antibacterial YR26 peptide bound to SDS micelle. (A) Superposition of backbone atoms of 20 lowest energy structures of YR26 calculated from CYANA. The pdb coordinates of the structures of YR26 have been deposited to pdb data bank, accession number 6A8Y. (B) Ribbon representation of a selected structure of YR26 showing a helix-turn-helix fold. The sidechains of N-terminal aromatic residues are shown as space-fill whereas sidechains of basic residues are shown as sticks. (C) The electrostatic surface potential surface of YR26 in two different orientations highlighting cationic patches.

Article Snippet: A synthetic gene of YR26, from Shanghai ShineGene Molecular Biotech, Inc, was cloned as a fusion protein in a pET14b vector containing hydrophobic KSI at the N-terminus.

Techniques:

Journal: ACS Omega

Article Title: Cell-Selective Pore Forming Antimicrobial Peptides of the Prodomain of Human Furin: A Conserved Aromatic/Cationic Sequence Mapping, Membrane Disruption, and Atomic-Resolution Structure and Dynamics

doi: 10.1021/acsomega.8b01876

Figure Lengend Snippet: Backbone motional characteristics of YR26 in SDS micelle. (A) Longitudinal ( R 1 ) relaxation rate of residues of YR26, with an average of 1.44 s –1 . (B) Transverse ( R 2 ) relaxation rate of residues of YR26, with an average of 10.93 s –1 . (C) Heteronuclear NOEs of residues of YR26, with an average of 0.47. The dotted line in the plot indicates average values of R 1 , R 2 , and heteronuclear NOE.

Article Snippet: A synthetic gene of YR26, from Shanghai ShineGene Molecular Biotech, Inc, was cloned as a fusion protein in a pET14b vector containing hydrophobic KSI at the N-terminus.

Techniques:

Journal: ACS Omega

Article Title: Cell-Selective Pore Forming Antimicrobial Peptides of the Prodomain of Human Furin: A Conserved Aromatic/Cationic Sequence Mapping, Membrane Disruption, and Atomic-Resolution Structure and Dynamics

doi: 10.1021/acsomega.8b01876

Figure Lengend Snippet: Minimum Inhibitory Concentration to Screen Various Prodomain Truncations a

Article Snippet: A synthetic gene of YR26, from Shanghai ShineGene Molecular Biotech, Inc, was cloned as a fusion protein in a pET14b vector containing hydrophobic KSI at the N-terminus.

Techniques: Concentration Assay

Journal: ACS Omega

Article Title: Cell-Selective Pore Forming Antimicrobial Peptides of the Prodomain of Human Furin: A Conserved Aromatic/Cationic Sequence Mapping, Membrane Disruption, and Atomic-Resolution Structure and Dynamics

doi: 10.1021/acsomega.8b01876

Figure Lengend Snippet: Minimum Inhibitory Concentration (MIC, in μM) of Deletion Analogs of YR26 a

Article Snippet: A synthetic gene of YR26, from Shanghai ShineGene Molecular Biotech, Inc, was cloned as a fusion protein in a pET14b vector containing hydrophobic KSI at the N-terminus.

Techniques: Concentration Assay

Journal: ACS Omega

Article Title: Cell-Selective Pore Forming Antimicrobial Peptides of the Prodomain of Human Furin: A Conserved Aromatic/Cationic Sequence Mapping, Membrane Disruption, and Atomic-Resolution Structure and Dynamics

doi: 10.1021/acsomega.8b01876

Figure Lengend Snippet: Thermodynamic Interactions between LPS and Prodomain Peptides Determined from ITC a

Article Snippet: A synthetic gene of YR26, from Shanghai ShineGene Molecular Biotech, Inc, was cloned as a fusion protein in a pET14b vector containing hydrophobic KSI at the N-terminus.

Techniques: