• Analytical Science and Technology
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• v.28 no.2
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• pp.106-111
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• 2015

Changes in antimicrobial peptide-lipid mixtures were investigated using ³¹P solid-state nuclear magnetic resonance spectroscopy. An antimicrobial peptide, protegrin-1, and phosphatidylcholine were deposited on a thin cover glass and incubated under a relative humidity of 95%. The changes in the mixtures were observed after hydration or air-drying. How repetitive hydration and drying changed the phase of the sample was also observed. The degrees of disruption of the well-aligned bilayers of phosphatidylcholine were determined quantitatively by simulating the experimental spectra. The peptide-lipid mixtures changed reversibly after hydration and drying, and the samples reached an equilibrium state after several repetitions.

• Analytical Science and Technology
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• v.26 no.1
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• pp.61-66
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• 2013

The activity of an antimicrobial peptide, protegrin-1, on the well-aligned lipid bilayer deposited on a thin coverglass plate was investigated by $^2H$ solid-state NMR spectroscopy. Orientational distribution and molecular motion in the lipid bilayer were determined from $^2H$ solid-state NMR spectrum. Reorientational motion of lipid molecules in the vacuum-dried state was found to be small but their orientational distribution was not able to be determined. As storage times were longer, the order of the alignment of lipid molecules in the lipid bilayer and percentages involved in the toroidal pore structures increased. We found that much longer time is required to get the equilibrium state of the peptide-lipid mixture under our experimental condition for investigating the action of the antimicrobial peptide like protegrin-1 on the lipid bilayers deposited on the thin coverglass plates.

• Bulletin of the Korean Chemical Society
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• v.31 no.2
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• pp.372-378
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• 2010

Membrane disruption by an antimicrobial peptide, protegrin-1 (PG-1), was investigated by measuring the $^2H$ solid-state nuclear magnetic resonance (SSNMR) spectra of 1-palmitoyl-$d_{31}$-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC_$d_{31}$) in the mixture of PG-1 and POPC_$d_{31}$ lipids deposited on thin cover-glass plates. The experimental line shapes of anisotropic $^2H$ SSNMR spectra measured at various peptide-to-lipid (P/L) ratios were simulated reasonably by assuming the mosaic spread of bilayers containing pore structures or the coexistence of the mosaic spread of bilayers and a fast-tumbling isotropic phase. Within a few days of incubation in the hydration chamber, the pores were formed by the peptide in the POPC_$d_{31}$ and POPC_$d_{31}$/cholesterol membranes. However, the formation of the pores was not clear in the POPC_$d_{31}$/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol (POPG) membrane. Over a hundred days after hydration, a rapidly rotating isotropic phase increased in the POPC_$d_{31}$ and the POPC_$d_{31}$/cholesterol membranes with the higher P/L ratios, but no isotropic phase appeared in the POPC_$d_{31}$/POPG membrane. Cholesterol added in the POPC bilayer acted as a stabilizer of the pore structure and suppressed the formation of a fast-tumbling isotropic phase.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.669-671
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• 2000

Antimicrobial cationic peptides have attracted increasing research and clinical interest as a natural antibiotics due to their broad spectrum of antimicrobial activites and the rapid development of multidrug-resistant pathogenic microorganisms. In this study, first, we synthesized artificial fusion partner and cationic peptide genes (lactoferricin, magainin, protegrin-1, and indolicidin). Second, we constructed recombinant expression vectors and then transformed Pichia pastoris. Finally, expressed cationic peptides were purified and tested for their antimicrobial activites. Antimicrobial activity has been tested upon the appearance of clearing zone on the plate with the lawn of gram negative E.coli XL- I blue and garm positive Staphylococcus aureus. Protegrin-1 and Indolicidin have apparant activity of cationic peotides. This fusion technique may lead to a general and suitable tool for production of pure antimicrobial cationic peptides in Pichia pastoris.

• Analytical Science and Technology
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• v.26 no.6
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• pp.395-400
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• 2013

The hydration and antimicrobial peptide effects on the lipid alignment on the surface of a thin glass plate were investigated by using the solid-state nuclear magnetic resonance spectroscopy. Pure 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphotidylcholine (POPC) lipid molecules were well aligned on the surface of a thin glass plate without direct hydration by placing the sample for a few days in the desiccator containing a saturated sodium phosphate dibasic solution, which provided 95% relative humidity. But there was a big difference between two lipid phases, the one of which was hydrated by placing the sample for a few days in the desiccator with 95% relative humidity without direct hydration and the other was directly hydrated by dropping water and then placed for a few days in the same desiccator. The surface mobility of POPC molecules in a lipid bilayer phase was much activated by water. While the time for the POPC to align on the glass plate surface was short, the time for the PG-1/POPC mixture to reach the its equilibrium state was long.

• Bulletin of the Korean Chemical Society
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• v.33 no.2
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• pp.426-432
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• 2012

The activity of an antimicrobial peptide, protegrin-1 (PG-1), on lipid membranes was investigated using solidstate NMR and a new sampling method that employed mechanically aligned bilayers between thin glass plates. At 95% hydration and full hydration, the peptide respectively disrupted 25% and 86% of the aligned 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphotidylcholine (POPC) bilayers at a P/L (peptide-to-lipid) ratio of 1/20 under the new experimental conditions. The kinetics of the POPC bilayers disruption appeared to be diffusioncontrolled. The presence of cholesterol at 95% hydration and full hydration reduced the peptide disruption of the aligned POPC bilayers to less than 10% and 35%, respectively. A comparison of the equilibrium states of heterogeneously and homogeneously mixed peptides and lipids demonstrated the importance of peptide binding to the biomembrane for whole membrane disruption.