Keun-Hyeung Lee

Effect of D-amino acid substitution on the stability, the secondary structure, and the activity of membrane-active peptide.

Several diastereomers and an enantiomer of KKVVFKVKFKK, an antimicrobial peptide that acts on the lipid membrane of pathogens were synthesized to investigate the effect of D-amino acid substitution on stability, secondary structure, and activity. The stability of the peptide in serum was improved greatly by the D-amino acid substitutions. D-Amino acid substitutions at the N- and/or C-terminal of the peptide, which had little effect on the alpha-helical structure, and all D-amino acid substitutions that formed a left-handed alpha-helix maintained antimicrobial activity, whereas D-amino acid substitutions in the middle of the amino acid sequence disrupted the alpha-helical structure, resulting in the complete loss of activity. This result confirmed that the peptide did not interact with chiral receptors, enzymes, or any chiral component of the membrane. D-Amino acid substitutions at the termini reduced the inhibition of the activity by heat-inactivated serum, which indicated that local change of chirality or change of secondary structure induced by D-amino acid substitutions might affect the interactions between the peptide and certain components in the serum. The present study suggests that partial D-amino acid substitution is a useful technique to improve the in vivo activity of antimicrobial peptides.

Selective and sensitive ratiometric detection of Hg(II) ions using a simple amino acid based sensor.

Synthesis of a novel pyrene derivative sensor (Py-Met) based on amino acid and its fluorescent behavior for Hg(II) in water was investigated. Upon Hg(II) binding, the Py-Met-bearing sulfonamide group exhibited a considerable excimer emission at 480 nm along with a decrease of monomer emission at 383 nm. Py-Met allows a selective and sensitive ratiometric detection of Hg(II) without any interference from other metal ions.

Turn-on Fluorescent Chemosensor Based on an Amino Acid for Pb(II) and Hg(II) Ions in Aqueous Solutions and Role of Tryptophan for Sensing

This communication presents a fluorescent chemosensor for detecting Pb(II) and Hg(II) in aqueous solutions. The sensor showed a turn-on response to Pb(II) by an enhancement of emission intensity at 380 nm and to Hg(II) by an enhancement of emission intensities at 380 and 475 nm. We have first characterized a unique function of tryptophan as a ligand as well as a quencher for recognition and fluorescent change by a metal binding event.

Selective and sensitive ratiometric detection of Hg2+ in 100% aqueous solution with triazole-based dansyl probe

An amino acid-based fluorescent sensor (3) bearing a triazole group was synthesized by solid phase synthesis using click chemistry. 3 showed highly sensitive ratiometric response to Hg2+ in 100% aqueous solution. The dissociation constant for Hg2+ in aqueous solution was calculated to be 672 nM. 3 exhibited selective response to Hg2+ among 13 tested metal ions and the sensitive detection of Hg2+ by 3 was not interfered by other heavy metal ions, Group I and II metal ions. 1H NMR analysis revealed that two sulfonamide and triazole groups of 3 played a critical role in the interactions with Hg2+. 3 penetrated live cells and detected intracellular Hg2+. 3 allowed selective and sensitive ratiometric detection of Hg2+ in live cells as well as in 100% aqueous solution.

A highly sensitive and selective detection of Hg(II) in 100% aqueous solution with fluorescent labeled dimerized Cys residues

A simple design of a ratiometric fluorescent sensor for detecting Hg(II) ion in 100% aqueous solution was demonstrated, based on the structure of dimerized Cys residues with two dansyl fluorophores. The sensor highly sensitively and selectively detected mercury ion (K(d) = 41 nM) in 100% aqueous solution via a turn-on and ratiometric response. The sensor showed no interferences of other metal ions and satisfied for monitoring the maximum allowable level (2 ppb) of mercury ion in drinking water demanded by EPA via a turn-on response.

Novel thiazolidinedione derivatives with anti-obesity effects: Dual action as PTP1B inhibitors and PPAR-γ activators

Benzylidene-2,4-thiazolidinedione derivatives with substitutions at both the ortho and para-positions of the phenyl group were synthesized as PTP1B inhibitors with IC(50) values in a low micromolar range. Compound 18l, the lowest, bore an IC(50) of 1.3 μM. In a peroxisome proliferator-activated receptor-γ (PPAR-γ) promoter reporter gene assay, 18l was found to activate the transcription of the reporter gene with potencies comparable to those of troglitazone, rosiglitazone, and pioglitazone. In vivo efficacy of 18l as an anti-obesity and hypoglycemic agent was evaluated in a mouse model system. Compound 18l significantly suppressed weight gain and significantly improved blood parameters such as TG, total cholesterol and NEFA without overt toxic effects.

Formylchromone derivatives as a novel class of protein tyrosine phosphatase 1B inhibitors

Formylchromone inhibits a human protein tyrosine phosphatase PTP1B with a IC(50) value of 73 microM. The chemical reactivity of formylchromone was adjusted by substitution at various positions of the formylchromone skeleton. In an initial assessment of the structure-activity relationship, the most potent inhibitor showed an IC(50) of 4.3 microM against PTP1B and strong or medium selectivity against other human PTPases, LAR and TC-PTP. This compound, however, was not selective against microbial PTPases, YPTP1 and YOP. The potency and selectivity of the formylchromone derivatives expecting further improvements provides a novel pharmacophore for the design of drugs for the treatment of type 2 diabetes and obesity.

Selective and Sensitive Detection of Heavy Metal Ions in 100% Aqueous Solution and Cells with a Fluorescence Chemosensor Based on Peptide Using Aggregation-Induced Emission

A fluorescent peptidyl chemosensor for the detection of heavy metal ions in aqueous solution as well as in cells was synthesized on the basis of the peptide receptor for the metal ions using an aggregation-induced emission fluorophore. The peptidyl chemosensor (1) bearing tetraphenylethylene fluorophore showed an exclusively selective turn-on response to Hg(2+) among 16 metal ions in aqueous buffered solution containing NaCl. The peptidyl chemosensor complexed Hg(2+) ions and then aggregated in aqueous buffered solution, resulting in the significant enhancement (OFF-On) of emissions at around 470 nm. The fluorescent sensor showed a highly sensitive response to Hg(2+), and about 1.0 equiv of Hg(2+) was enough for the saturation of the emission intensity change. The detection limit (5.3 nM, R(2) = 0.99) of 1 for Hg(2+) ions was lower than the maximum allowable level of Hg(2+) in drinking water by EPA. Moreover, the peptidyl chemosensor penetrated live cells and detected intracellular Hg(2+) ions by the turn-on response.

The effect of charge increase on the specificity and activity of a short antimicrobial peptide

By using short linear antimicrobial peptides as a model system, the effect of peptide charge on the specificity between Candida albicans (fungi) and Gram-positive bacteria was investigated. In a present study, we added and/or deleted lysine residue(s) at the C-terminal and/or N-terminal end(s) of an antimicrobial peptide (KKVVFKVKFK-NH(2)) and synthesized the peptides that had similar alpha helical structures in a lipid membrane mimic condition. The increase of peptide charge improved antifungal activity without the change of antibacterial activity. Structure-activity relationship study about the peptides revealed that the net positive charge must play an important role in the specificity between C. albicans and Gram-positive bacteria and the increase of the net positive charge without the moderate change of secondary structure could improve activity for C. albicans rather than Gram-positive bacteria.

Highly selectively monitoring heavy and transition metal ions by a fluorescent sensor based on dipeptide

Fluorescent sensor (DMH) based on dipeptide was efficiently synthesized in solid phase synthesis. The dipeptide sensor shows sensitive response to Ag(I), Hg(II), and Cu(II) among 14 metal ions in 100% aqueous solution. The fluorescent sensor differentiates three heavy metal ions by response type; turn on response to Ag(I), ratiometric response to Hg(II), and turn off detection of Cu(II). The detection limits of the sensor for Ag(I) and Cu(II) were much lower than the EPAs drinking water maximum contaminant levels (MCL). Specially, DMH penetrated live cells and detected intracellular Ag(+) by turn on response. We described the fluorescent change, binding affinity, detection limit for the metal ions. The study of a heavy metal-responsive sensor based on dipeptide demonstrates its potential utility in the environment field.