Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 23
Filter
Add more filters










Publication year range
1.
Front Cell Dev Biol ; 4: 86, 2016.
Article in English | MEDLINE | ID: mdl-27617260

ABSTRACT

RWRWRW-NH2 (MP196) is an amphipathic hexapeptide that targets the bacterial cytoplasmic membrane and inhibits cellular respiration and cell wall synthesis. In previous studies it showed promising activity against Gram-positive bacteria and no significant cytotoxicity or hemolysis. MP196 is therefore used as lead structure for developing more potent antibiotic derivatives. Here we present a more comprehensive study on the parent peptide MP196 with regard to clinically relevant parameters. We found that MP196 acts rapidly bactericidal killing 97% of initial CFU within 10 min at two times MIC. We were unable to detect resistance in standard 24 and 48 h resistance frequency assays. However, MP196 was effective against some but not all MRSA and VISA strains. Serum binding of MP196 was intermediate and we confirmed its low toxicity against mammalian cell lines. MP196 did neither induce NFκB activation nor cause an increase in IL8 levels at 250 µg/mL, and no IgE-dependent activation of basophil granulocytes was detected at 500 µg/mL. Yet, MP196 demonstrated acute toxicity in mice upon injection into the blood stream. Phase contrast microscopy of mouse blood treated with MP196 revealed a shrinking of erythrocytes at 250 µg/mL and severe morphological changes and lysis of erythrocytes at 500 µg/mL. These data suggest that MP196 derivatization directed at further lowering hemolysis could be instrumental in overcoming acute toxicity. The assessment of hemolysis is a critical step in the evaluation of the clinical potential of promising antimicrobial peptides and should be accompanied by microscopy-based morphological analysis of blood cells.

2.
Chem Commun (Camb) ; 52(32): 5561-4, 2016 Apr 25.
Article in English | MEDLINE | ID: mdl-27020540

ABSTRACT

Lipidated DNAzymes or a lipidated Cu(II)-complex and lipidated aptamer sequences form supramolecular assemblies of micellar nucleoapzymes for the enhanced oxidation of dopamine to aminochrome. The catalytic functions of the micellar nucleoapzymes are attributed to the concentration of the substrate, using the aptamer units, in close proximity to the active sites.


Subject(s)
Dopamine/chemistry , Indolequinones/chemistry , Micelles , Catalysis , Oxidation-Reduction
3.
Biochim Biophys Acta ; 1858(5): 1004-11, 2016 May.
Article in English | MEDLINE | ID: mdl-26603779

ABSTRACT

Antimicrobial peptides are a potent class of antibiotics. In the Gram-positive model organism Bacillus subtilis the synthetic peptide RWRWRW-NH2 integrates into the bacterial membrane and delocalizes essential peripheral membrane proteins involved in cell wall biosynthesis and respiration. A lysine residue has been added to the hexapeptide core structure, either C or N-terminally. Lipidation of the lysine residues by a C8-acyl chain significantly improved antibacterial activity against both Gram-positive and Gram-negative bacteria. Here, we report a comparative proteomic study in B. subtilis on the mechanism of action of the lipidated and non-lipidated peptides. All derivatives depolarized the bacterial membrane without forming pores and all affected cell wall integrity. Proteomic profiling of the bacterial stress responses to the small RW-rich antimicrobial peptides was reflective of non-disruptive membrane integration. Overall, our results indicate that antimicrobial peptides can be derivatized with lipid chains enhancing antibacterial activity without significantly altering the mechanism of action. This article is part of a Special Issue entitled: Antimicrobial peptides edited by Karl Lohner and Kai Hilpert.


Subject(s)
Anti-Bacterial Agents/pharmacology , Antimicrobial Cationic Peptides/pharmacology , Bacillus subtilis/drug effects , Cell Membrane/drug effects , Cell Wall/drug effects , Escherichia coli/drug effects , Gene Expression Regulation, Bacterial , Amino Acid Sequence , Anti-Bacterial Agents/chemistry , Antimicrobial Cationic Peptides/chemical synthesis , Antimicrobial Cationic Peptides/chemistry , Arginine/chemistry , Arginine/metabolism , Bacillus subtilis/chemistry , Bacillus subtilis/genetics , Bacillus subtilis/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Cell Membrane/chemistry , Cell Membrane/metabolism , Cell Wall/chemistry , Cell Wall/metabolism , Drug Resistance, Multiple, Bacterial/drug effects , Escherichia coli/chemistry , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression Profiling , Microbial Sensitivity Tests , Molecular Sequence Data , Structure-Activity Relationship , Tryptophan/chemistry , Tryptophan/metabolism
4.
J Am Chem Soc ; 138(1): 164-72, 2016 Jan 13.
Article in English | MEDLINE | ID: mdl-26652164

ABSTRACT

A novel concept to improve the catalytic functions of nucleic acids (DNAzymes) is introduced. The method involves the conjugation of a DNA recognition sequence (aptamer) to the catalytic DNAzyme, yielding a hybrid structure termed "nucleoapzyme". Concentrating the substrate within the "nucleoapzyme" leads to enhanced catalytic activity, displaying saturation kinetics. Different conjugation modes of the aptamer/DNAzyme units and the availability of different aptamer sequences for a substrate provide diverse means to design improved catalysts. This is exemplified with (i) The H2O2-mediated oxidation of dopamine to aminochrome using a series of hemin/G-quadruplex-dopamine aptamer nucleoapzymes. All nucleoapzymes reveal enhanced catalytic activities as compared to the separated DNAzyme/aptamer units, and the most active nucleoapzyme reveals a 20-fold enhanced activity. Molecular dynamics simulations provide rational assessment of the activity of the various nucleoapzymes. The hemin/G-quadruplex-aptamer nucleoapzyme also stimulates the chiroselective oxidation of L- vs D-DOPA by H2O2. (ii) The H2O2-mediated oxidation of N-hydroxy-L-arginine to L-citrulline by a series of hemin/G-quadruplex-arginine aptamer conjugated nucleoapzymes.


Subject(s)
Aptamers, Nucleotide/chemistry , DNA, Catalytic/chemistry , G-Quadruplexes , Hemin/chemistry , Binding Sites , Catalysis
5.
Chem Sci ; 7(5): 3092-3101, 2016 May 01.
Article in English | MEDLINE | ID: mdl-29997801

ABSTRACT

The rational design of a set of hemin/G-quadruplex (hGQ)-dopamine binding aptamer (DBA) conjugates, acting as nucleoapzymes, is described. The nucleoapzyme constructs consist of a hGQ DNAzyme as a catalytic unit and DBA as a substrate binding unit that are brought into spatial proximity by a duplex scaffold composed of complementary oligonucleotide strands. When the hGQ unit is linked to the duplex scaffold via a single-strand DNA tether of variable length, the resulting nucleoapzymes reveal a moderate catalytic enhancement toward the H2O2-mediated oxidation of dopamine to aminochrome as compared to the process stimulated by the separated hGQ and DBA units (5-7 fold enhancement). This limited enhancement is attributed to inappropriate spatial positioning of the hGQ in respect to the dopamine binding site, and/or to the flexibility of the tether that links the hGQ catalytic site to the double-stranded scaffold. To solve this, rigidification of the hGQ/DBA conjugates by triplex oligonucleotide structures that anchor the hGQ to a duplex domain associated with the DBA units was achieved. By the sequential, programmed, triplex-controlled rigidification of the hGQ/DBA structure, a nucleoapzyme with superior catalytic activity toward the oxidation of dopamine to aminochrome is identified (30-fold catalytic enhancement). Molecular dynamics simulations reveal that in the resulting highly active rigidified nucleoapzyme structure, the hGQ catalytic site is positioned in spatial proximity to the opening of the DBA substrate binding site, thus rationalizing and supporting the enhanced catalytic functions of the system. Finally, the most active nucleoapzyme system was subjected to fuel- and anti-fuel strands that separate and re-assemble the nucleoapzyme structure, allowing "ON" and "OFF" switching of the nucleoapzyme catalytic functions.

6.
Angew Chem Int Ed Engl ; 54(40): 11652-6, 2015 Sep 28.
Article in English | MEDLINE | ID: mdl-25959900

ABSTRACT

DNAzyme-capped mesoporous SiO2 nanoparticles (MP SiO2 NPs) are applied as stimuli-responsive containers for programmed synthesis. Three types of MP SiO2 NPs are prepared by loading the NPs with Cy3-DBCO (DBCO=dibenzocyclooctyl), Cy5-N3 , and Cy7-N3 , and capping the NP containers with the Mg(2+) , Zn(2+) , and histidine-dependent DNAzyme sequences, respectively. In the presence of Mg(2+) and Zn(2+) ions as triggers, the respective DNAzyme-capped NPs are unlocked, leading to the "click" reaction product Cy3-Cy5. In turn, in the presence of Mg(2+) ions and histidine as triggers the second set of DNAzyme-capped NPs is unlocked leading to the Cy3-Cy7 conjugated product. The unloading of the respective NPs and the time-dependent formation of the products are followed by fluorescence spectroscopy (FRET). A detailed kinetic model for the formation of the different products is formulated and it correlates nicely with the experimental results.


Subject(s)
DNA, Catalytic/metabolism , Nanoparticles/chemistry , Nanoparticles/metabolism , Silicon Dioxide/chemistry , Silicon Dioxide/metabolism , Kinetics , Particle Size , Porosity , Spectrometry, Fluorescence , Surface Properties
7.
J Comput Aided Mol Des ; 29(7): 643-54, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25877490

ABSTRACT

The binding properties of sequence-specific nucleic acids (aptamers) to low-molecular-weight ligands, macromolecules and even cells attract substantial scientific interest. These ligand-DNA complexes found different applications for sensing, nanomedicine, and DNA nanotechnology. Structural information on the aptamer-ligand complexes is, however, scarce, even though it would open-up the possibilities to design novel features in the complexes. In the present study we apply molecular docking simulations to probe the features of an experimentally documented L-argininamide aptamer complex. The docking simulations were performed using AutoDock 4.0 and YASARA Structure software, a well-suited program for following intermolecular interactions and structures of biomolecules, including DNA. We explored the binding features of a DNA aptamer to L-argininamide and to a series of arginine derivatives or arginine-like ligands. We find that the best docking results are obtained after an energy-minimization of the parent ligand-aptamer complexes. The calculated binding energies of all mono-substituted guanidine-containing ligands show a good correlation with the experimentally determined binding constants. The results provide valuable guidelines for the application of docking simulations for the prediction of aptamer-ligand structures, and for the design of novel features of ligand-aptamer complexes.


Subject(s)
Aptamers, Nucleotide/chemistry , Aptamers, Nucleotide/metabolism , Arginine/analogs & derivatives , Arginine/chemistry , Arginine/metabolism , Computer Simulation , Ligands , Magnetic Resonance Spectroscopy , Molecular Docking Simulation , Software
8.
Chem Commun (Camb) ; 51(6): 1100-3, 2015 Jan 21.
Article in English | MEDLINE | ID: mdl-25449885

ABSTRACT

Two-sized luminescent nucleic acid-functionalized Ag nanoclusters (NCs) are implemented for the analysis and multiplexed detection of adenosine monophosphate, AMP, and of cocaine using aptamer-ligand complexes.


Subject(s)
Aptamers, Nucleotide/chemistry , Chemistry Techniques, Analytical , Nanoparticles/chemistry , Silver/chemistry , Adenosine Monophosphate/analysis , Cocaine/analysis , Ligands , Luminescence
9.
ACS Nano ; 8(11): 11666-73, 2014 Nov 25.
Article in English | MEDLINE | ID: mdl-25327411

ABSTRACT

Luminescent nucleic acid-stabilized Ag nanoclusters (Ag NCs) are applied for the optical detection of DNA and for the multiplexed analysis of genes. Two different sensing modules including Ag NCs as luminescence labels are described. One sensing module involves the assembly of a three-component sensing module composed of a nucleic acid-stabilized Ag NC and a quencher-modified nucleic acid hybridized with a nucleic acid scaffold that is complementary to the target DNA. The luminescence of the Ag NCs is quenched in the sensing module nanostructure. The strand displacement of the scaffold by the target DNA separates the nucleic acid-functionalized Ag NCs, leading to the turned-on luminescence of the NCs and to the optical readout of the sensing process. By implementing two different-sized Ag NC-modified sensing modules, the parallel multiplexed analysis of two genes (the Werner Syndrome gene and the HIV, human immunodeficiency, gene), using 615 and 560 nm luminescent Ag NCs, is demonstrated. The second sensing module includes the nucleic acid functionalized Ag NCs and the quencher-modified nucleic acid hybridized with a hairpin DNA scaffold. The luminescence of the Ag NCs is quenched in the sensing module. Opening of the hairpin by the target DNA triggers the luminescence of the Ag NCs, due to the spatial separation of the Ag NCs/quencher units. The system is applied for the optical detection of the BRAC1 gene. In addition, by implementing two-sized Ag NCs, the multiplexed analysis of two genes by the hairpin sensing module approach is demonstrated.


Subject(s)
DNA/genetics , Nanostructures , Nucleic Acids/chemistry , Quantum Dots , Silver/chemistry , Base Sequence , DNA Primers , Genes, BRCA1 , Humans
10.
Angew Chem Int Ed Engl ; 53(27): 7058-62, 2014 Jul 01.
Article in English | MEDLINE | ID: mdl-24838592

ABSTRACT

A mixed-linker solid-solution approach was employed to modify the metal sites and introduce structural defects into the mixed-valence Ru(II/III) structural analogue of the well-known MOF family [M3(II,II)(btc)2] (M=Cu, Mo, Cr, Ni, Zn; btc=benzene-1,3,5-tricarboxylate), with partly missing carboxylate ligators at the Ru2 paddle-wheels. Incorporation of pyridine-3,5-dicarboxylate (pydc), which is the same size as btc but carries lower charge, as a second, defective linker has led to the mixed-linker isoreticular derivatives of Ru-MOF, which display characteristics unlike those of the defect-free framework. Along with the creation of additional coordinatively unsaturated sites, the incorporation of pydc induces the partial reduction of ruthenium. Accordingly, the modified Ru sites are responsible for the activity of the "defective" variants in the dissociative chemisorption of CO2, the enhanced performance in CO sorption, the formation of hydride species, and the catalytic hydrogenation of olefins.


Subject(s)
Organic Chemicals/chemistry , Ruthenium/chemistry , Carbon Dioxide/chemistry , Carbon Monoxide/chemistry , Catalysis , Coordination Complexes/chemistry , Hydrogenation , Oxidation-Reduction
11.
Proc Natl Acad Sci U S A ; 111(14): E1409-18, 2014 Apr 08.
Article in English | MEDLINE | ID: mdl-24706874

ABSTRACT

Short antimicrobial peptides rich in arginine (R) and tryptophan (W) interact with membranes. To learn how this interaction leads to bacterial death, we characterized the effects of the minimal pharmacophore RWRWRW-NH2. A ruthenium-substituted derivative of this peptide localized to the membrane in vivo, and the peptide also integrated readily into mixed phospholipid bilayers that resemble Gram-positive membranes. Proteome and Western blot analyses showed that integration of the peptide caused delocalization of peripheral membrane proteins essential for respiration and cell-wall biosynthesis, limiting cellular energy and undermining cell-wall integrity. This delocalization phenomenon also was observed with the cyclic peptide gramicidin S, indicating the generality of the mechanism. Exogenous glutamate increases tolerance to the peptide, indicating that osmotic destabilization also contributes to antibacterial efficacy. Bacillus subtilis responds to peptide stress by releasing osmoprotective amino acids, in part via mechanosensitive channels. This response is triggered by membrane-targeting bacteriolytic peptides of different structural classes as well as by hypoosmotic conditions.


Subject(s)
Antimicrobial Cationic Peptides/metabolism , Membrane Proteins/metabolism , Bacillus subtilis/metabolism , Binding Sites , Cytochromes c/metabolism , Homeostasis , Lipid Bilayers , Phospholipids/metabolism
12.
ACS Comb Sci ; 15(11): 585-92, 2013 Nov 11.
Article in English | MEDLINE | ID: mdl-24147906

ABSTRACT

High systemic toxicity of antimicrobial peptides (AMPs) limits their clinical application to the treatment of topical infections; in parenteral systemic application of AMPs the problem of hemolysis is one of the first to be tackled. We now show that the selectivity of lipidated short synthetic AMPs can be optimized substantially by reducing their hemolytic activity without affecting their activity against methicillin resistant Staphylococcus aureus (MRSA). In order to identify the optimized peptides, two sets of 32 diastereomeric H-(D)Arg-WRWRW-(L)Lys(C(O)CnH2n+1)-NH2 (n = 7 or 9) peptides were prepared using a split-split procedure to perform a systematic L-to-D exchange scan on the central WRWRW-fragment. Compared to the all-L C8-lipidated lead sequence, diastereomeric peptides had very similar antibacterial properties, but were over 30 times less hemolytic. We show that the observed hemolysis and antibacterial activity is affected by both differences in lipophilicity of the different peptides and specific combinations of L- and D-amino acid residues. This study identified several peptides that can be used as tools to precisely unravel the origin of hemolysis and thus help to design even further optimized nontoxic very active short antibacterial peptides.


Subject(s)
Amino Acids/chemistry , Anti-Bacterial Agents/pharmacology , Erythrocytes/drug effects , Hemolysis/drug effects , Peptides/pharmacology , Acinetobacter baumannii/drug effects , Acinetobacter baumannii/growth & development , Anti-Bacterial Agents/chemical synthesis , Anti-Bacterial Agents/chemistry , Bacillus subtilis/drug effects , Bacillus subtilis/growth & development , Dose-Response Relationship, Drug , Escherichia coli/drug effects , Escherichia coli/growth & development , Humans , Microbial Sensitivity Tests , Peptides/chemical synthesis , Peptides/chemistry , Pseudomonas aeruginosa/drug effects , Staphylococcus aureus/drug effects , Staphylococcus aureus/growth & development , Stereoisomerism , Structure-Activity Relationship
13.
Dalton Trans ; 42(27): 9799-802, 2013 Jul 21.
Article in English | MEDLINE | ID: mdl-23525213

ABSTRACT

Binding of Leu-enkephalin and [Rh(III)(η(5)-Cp*)(η(6)-Tyr(1))]Leu-enkephalin to the recently published crystal structures of the µ- and δ-opioid receptor is studied. Docking of free Leu-enkephalin reveals two preferred conformations, one of which suggests an alternative binding site for the tyrosine residue. Furthermore, the three-dimensional solution structure of [Rh(III)(η(5)-Cp*)(η(6)-Tyr(1))]Leu-enkephalin was solved by using 2D NMR spectroscopic techniques.


Subject(s)
Enkephalin, Leucine/chemistry , Organometallic Compounds/chemistry , Receptors, Opioid/chemistry , Rhodium/chemistry , Binding Sites , Crystallography, X-Ray , Magnetic Resonance Spectroscopy , Models, Molecular , Molecular Structure
14.
Org Lett ; 15(12): 3126-9, 2013 Jun 21.
Article in English | MEDLINE | ID: mdl-24490777

ABSTRACT

A new Silyl-based Alkyne Modifying (SAM)-linker for the synthesis of C-terminal acetylene-derivatized peptides is reported. The broad scope of this SAM2-linker is illustrated by manual synthesis of peptides that are side-chain protected, fully deprotected, and disulfide-bridged. Synthesis of a 14-meric (KLAKLAK)2 derivative by microwave-assisted automated SPPS and a one-pot cleavage click procedure yielding protected 1,2,3-triazole peptide conjugates are also described.


Subject(s)
Acetylene/chemistry , Alkynes/chemistry , Immobilized Proteins/chemistry , Peptides/chemical synthesis , Silanes/chemistry , Acetylene/chemical synthesis , Amino Acid Sequence , Microwaves , Molecular Structure , Peptides/chemistry
15.
Beilstein J Org Chem ; 8: 1753-64, 2012.
Article in English | MEDLINE | ID: mdl-23209509

ABSTRACT

A series of small synthetic arginine and tryptophan containing peptides was prepared and analyzed for their antibacterial activity. The effect of N-terminal substitution with metallocenoyl groups such as ferrocene (FcCO) and ruthenocene (RcCO) was investigated. Antibacterial activity in different media, growth inhibition, and killing kinetics of the most active peptides were determined. The toxicity of selected derivatives was determined against erythrocytes and three human cancer cell lines. It was shown that the replacement of an N-terminal arginine residue with a metallocenoyl moiety modulates the activity of WRWRW-peptides against Gram-positive and Gram-negative bacteria. MIC values of 2-6 µM for RcCO-W(RW)(2) and 1-11 µM for (RW)(3) were determined. Interestingly, W(RW)(2)-peptides derivatized with ferrocene were significantly less active than those derivatized with ruthenocene which have similar structural but different electronic properties, suggesting a major influence of the latter. The high activities observed for the RcCO-W(RW)(2)- and (RW)(3)-peptides led to an investigation of the origin of activity of these peptides using several important activity-related parameters. Firstly, killing kinetics of the RcCO-W(RW)(2)-peptide versus killing kinetics of the (RW)(3) derivative showed faster reduction of the colony forming units for the RcCO-W(RW)(2)-peptide, although MIC values indicated higher activity for the (RW)(3)-peptide. This was confirmed by growth inhibition studies. Secondly, hemolysis studies revealed that both peptides did not lead to significant destruction of erythrocytes, even up to 500 µg/mL for (RW)(3) and 250 µg/mL for RcCO-W(RW)(2). In addition, toxicity against three human cancer cell lines (HepG2, HT29, MCF7) showed that the (RW)(3)-peptide had an IC(50) value of ~140 µM and the RcW(RW)(2) one of ~90 µM, indicating a potentially interesting therapeutic window. Both the killing kinetics and growth inhibition studies presented in this work point to a membrane-based mode of action for these two peptides, each having different kinetic parameters.

16.
J Org Chem ; 77(22): 9954-8, 2012 Nov 16.
Article in English | MEDLINE | ID: mdl-23116417

ABSTRACT

A novel linker for the synthesis of C-terminal acetylene-functionalized protected peptides is described. This SAM1 linker is applied in the manual Fmoc-based solid-phase peptide synthesis of Leu-enkephalin and in microwave-assisted automated synthesis of Maculatin 2.1, an antibacterial peptide that contains 18 amino acid residues. For the cleavage, treatment with tetramethylammonium fluoride results in protected acetylene-derivatized peptides. Alternatively, a one-pot cleavage-click procedure affords the protected 1,2,3-triazole conjugate in high yields after purification.


Subject(s)
Acetylene/chemistry , Acetylene/chemical synthesis , Alkynes/chemistry , Enkephalin, Leucine/chemistry , Enkephalin, Leucine/chemical synthesis , Peptides/chemistry , Peptides/chemical synthesis , Silanes/chemistry , Microwaves , Molecular Structure , Solid-Phase Synthesis Techniques
17.
Antimicrob Agents Chemother ; 56(11): 5749-57, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22926563

ABSTRACT

Mersacidin, gallidermin, and nisin are lantibiotics, antimicrobial peptides containing lanthionine. They show potent antibacterial activity. All three interfere with cell wall biosynthesis by binding lipid II, but they display different levels of interaction with the cytoplasmic membrane. On one end of the spectrum, mersacidin interferes with cell wall biosynthesis by binding lipid II without integrating into bacterial membranes. On the other end of the spectrum, nisin readily integrates into membranes, where it forms large pores. It destroys the membrane potential and causes leakage of nutrients and ions. Gallidermin, in an intermediate position, also readily integrates into membranes. However, pore formation occurs only in some bacteria and depends on membrane composition. In this study, we investigated the impact of nisin, gallidermin, and mersacidin on cell wall integrity, membrane pore formation, and membrane depolarization in Bacillus subtilis. The impact of the lantibiotics on the cell envelope was correlated to the proteomic response they elicit in B. subtilis. By drawing on a proteomic response library, including other envelope-targeting antibiotics such as bacitracin, vancomycin, gramicidin S, or valinomycin, YtrE could be identified as the most reliable marker protein for interfering with membrane-bound steps of cell wall biosynthesis. NadE and PspA were identified as markers for antibiotics interacting with the cytoplasmic membrane.


Subject(s)
Bacillus subtilis/drug effects , Bacterial Proteins/biosynthesis , Bacteriocins/pharmacology , Cell Membrane/drug effects , Cell Wall/drug effects , Nisin/pharmacology , Peptides/pharmacology , Bacillus subtilis/genetics , Bacillus subtilis/metabolism , Bacterial Proteins/antagonists & inhibitors , Bacterial Proteins/genetics , Biological Transport/drug effects , Biomarkers/metabolism , Cell Membrane/genetics , Cell Membrane/metabolism , Cell Wall/genetics , Cell Wall/metabolism , Electrophoresis, Gel, Two-Dimensional , Membrane Potentials/drug effects , Potassium/metabolism , Proteome/antagonists & inhibitors , Proteome/genetics , Proteome/metabolism , Structure-Activity Relationship , Uridine Diphosphate N-Acetylmuramic Acid/analogs & derivatives , Uridine Diphosphate N-Acetylmuramic Acid/metabolism
18.
J Am Chem Soc ; 134(25): 10321-4, 2012 Jun 27.
Article in English | MEDLINE | ID: mdl-22671299

ABSTRACT

The bioconjugation of organometallic complexes with peptides has proven to be a novel approach for drug discovery. We report the facile and chemoselective reaction of tyrosine-containing G-protein-coupled receptor (GPCR) peptides with [Cp*Rh(H(2)O)(3)](OTf)(2), in water, at room temperature, and at pH 5-6. We have focused on three important GPCR peptides; namely, [Tyr(1)]-leu-enkephalin, [Tyr(4)]-neurotensin(8-13), and [Tyr(3)]-octreotide, each of which has a different position for the tyrosine residue, together with competing functionalities. Importantly, all other functional groups present, i.e., amino, carboxyl, disulfide, phenyl, and indole, were not prominent sites of reactivity by the Cp*Rh tris aqua complex. Furthermore, the influence of the Cp*Rh moiety on the structure of [Tyr(3)]-octreotide was characterized by 2D NMR, resulting in the first representative structure of an organometallic-peptide complex. The biological consequences of these Cp*Rh-peptide complexes, with respect to GPCR binding and growth inhibition of MCF7 and HT29 cancer cells, will be presented for [(η(6)-Cp*Rh-Tyr(1))-leu-enkephalin](OTf)(2) and [(η(6)-Cp*Rh-Tyr(3))-octreotide](OTf)(2).


Subject(s)
Models, Molecular , Organometallic Compounds/chemistry , Peptides/chemistry , Receptors, G-Protein-Coupled/chemistry , Rhodium/chemistry , Tyrosine/chemistry , Binding, Competitive , Breast Neoplasms/drug therapy , Female , HT29 Cells , Humans , Inhibitory Concentration 50 , Magnetic Resonance Spectroscopy , Organometallic Compounds/pharmacology , Protein Binding/drug effects , Protein Structure, Tertiary , Receptors, G-Protein-Coupled/metabolism
19.
J Med Chem ; 55(3): 1296-302, 2012 Feb 09.
Article in English | MEDLINE | ID: mdl-22243686

ABSTRACT

The activity of antimicrobial peptides (AMPs) that contain a large proportion of histidine residues (pK(a) ∼ 6) depends on the physiological pH environment. Advantages of these AMPs include high activity in slightly acidic areas of the human body and relatively low toxicity in other areas. Also, many AMPs are highly active in a multivalent form, but this often increases toxicity. Here we designed pH dependent amphiphilic compounds consisting of multiple ultrashort histidine lipopeptides on a triazacyclophane scaffold, which showed high activity toward Aspergillus fumigatus and Cryptococcus neoformans at acidic pH, yet remained nontoxic. In vivo, treatment with a myristic acid conjugated trivalent histidine-histidine dipeptide resulted in 55% survival of mice (n = 9) in an otherwise lethal murine lung Aspergillus infection model. Fungal burden was assessed and showed completely sterile lungs in 80% of the mice (n = 5). At pH 5.5 and 7.5, differing peptide-membrane interactions and peptide nanostructures were observed. This study underscores the potential of unique AMPs to become the next generation of clinical antimicrobial therapy.


Subject(s)
Antifungal Agents/chemical synthesis , Antimicrobial Cationic Peptides/chemical synthesis , Lipopeptides/chemical synthesis , Animals , Anti-Bacterial Agents/chemical synthesis , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Antifungal Agents/chemistry , Antifungal Agents/pharmacology , Antimicrobial Cationic Peptides/chemistry , Antimicrobial Cationic Peptides/pharmacology , Aspergillus fumigatus/drug effects , Candida albicans/drug effects , Cell Line , Cryptococcus neoformans/drug effects , Female , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Hemolysis , Humans , Hydrogen-Ion Concentration , Lipopeptides/chemistry , Lipopeptides/pharmacology , Lung/microbiology , Mice , Mice, Inbred ICR , Microbial Sensitivity Tests , Nanostructures , Polymers , Pulmonary Aspergillosis/drug therapy , Pulmonary Aspergillosis/microbiology , Structure-Activity Relationship
20.
Org Biomol Chem ; 10(5): 1088-92, 2012 Feb 07.
Article in English | MEDLINE | ID: mdl-22179680

ABSTRACT

We describe the synthesis and coordination behaviour to copper(II) of two close structural triazacyclophane-based mimics of two often encountered aspartic acid and histidine containing metalloenzyme active sites. Coordination of these mimics to copper(I) and their reaction with molecular oxygen leads to the formation of dimeric bis(µ-hydroxo) dicopper(II) complexes.


Subject(s)
Aspartic Acid/chemistry , Aza Compounds/chemistry , Biomimetic Materials/chemistry , Copper/chemistry , Heterocyclic Compounds, 2-Ring/chemistry , Histidine/chemistry , Metalloproteins/chemistry , Aspartic Acid/metabolism , Aza Compounds/metabolism , Biomimetic Materials/metabolism , Catalytic Domain , Coordination Complexes/chemistry , Coordination Complexes/metabolism , Copper/metabolism , Heterocyclic Compounds, 2-Ring/metabolism , Histidine/metabolism , Metalloproteins/metabolism , Oxygen/metabolism
SELECTION OF CITATIONS
SEARCH DETAIL
...