Enzyme catalysis prior to aromatic residues: Reverse engineering of a dephospho-CoA kinase.

The wide variety of protein structures and functions results from the diverse properties of the 20 canonical amino acids. The generally accepted hypothesis is that early protein evolution was associated with enrichment of a primordial alphabet, thereby enabling increased protein catalytic efficiencies and functional diversification. Aromatic amino acids were likely among the last additions to genetic code.

Identification of Protein Targets of Bioactive Small Molecules Using Randomly Photomodified Probes.

Identifying protein targets of bioactive small molecules often requires complex, lengthy development of affinity probes. We present a method for stochastic modification of small molecules of interest with a photoactivatable phenyldiazirine linker. The resulting isomeric mixture is conjugated to a hydrophilic copolymer decorated with biotin and a fluorophore.

Transition metal complexes bearing NHC ligands substituted with secondary polyfluoroalkyl groups.

Using three different approaches, racemic 1-(perfluoroalkyl)ethylamines were synthesized from perfluoroalkyl iodides or perfluoroalkanoic acids, and further transformed to the corresponding N,N'-disubstituted ethane-1,2-diimines and ethane-1,2-diamines as mixtures of diastereoisomers. Their cyclization afforded imidazolium or dihydroimidazolium salts, which led to silver or palladium complexes bearing NHC ligands substituted with secondary polyfluoroalkyl groups. The palladium complexes bearing a throwaway 3-chloropyridine ligand proved to be moderately active in the model Suzuki-Miyaura coupling.

LC-NMR Technique in the Analysis of Phytosterols in Natural Extracts.

The ability of LC-NMR to detect simultaneously free and conjugated phytosterols in natural extracts was tested. The advantages and disadvantages of a gradient HPLC-NMR method were compared to the fast composition screening using SEC-NMR method. Fractions of free and conjugated phytosterols were isolated and analyzed by isocratic HPLC-NMR methods.

Synthesis of alkylcarbonate analogs of O-acetyl-ADP-ribose.

The non-hydrolyzable alkylcarbonate analogs of O-acetyl-ADP-ribose have been synthesized from the phosphorylated ribose derivatives after coupling with AMP morpholidate promoted by mechanical grinding. The analogs were assessed for their ability to inhibit the human sirtuin homolog SIRT1.

Effect of phosphorus availability on the selection of species with different ploidy levels and genome sizes in a long-term grassland fertilization experiment.

Polyploidy and increased genome size are hypothesized to increase organismal nutrient demands, namely of phosphorus (P), which is an essential and abundant component of nucleic acids. Therefore, polyploids and plants with larger genomes are expected to be selectively disadvantaged in P-limited environments. However, this hypothesis has yet to be experimentally tested.

Quinacrine reactivity with prion proteins and prion-derived peptides.

Quinacrine is a drug that is known to heal neuronal cell culture infected with prions, which are the causative agents of neurodegenerative diseases called transmissible spongiform encephalopathies. However, the drug fails when it is applied in vivo. In this work, we analyzed the reason for this failure.

Compound instability in dimethyl sulphoxide, case studies with 5-aminopyrimidines and the implications for compound storage and screening.

The oxidation reactions of 5-aminopyrimidine derivatives in dimethyl sulphoxide (DMSO) were studied. The DMSO solutions of the studied compounds became deeply coloured within a few hours or days. The oxidation products can undergo further condensation reactions with the starting pyrimidines to yield bipyrimidines and/or pyrimidopteridines.

Identification by GC-EAD of the two-component trail-following pheromone of Prorhinotermes simplex (Isoptera, Rhinotermitidae, Prorhinotermitinae).

GC/MS analysis confirmed that neocembrene is the major component of the trail pheromone in the three species of the termite genus Prorhinotermes (P. simplex, P. canalifrons, P.

Cytochromes P450 reconstituted with NADPH: P450 reductase mimic the activating and detoxicating metabolism of the anticancer drug ellipticine in microsomes.

Objectives: Ellipticine is a potent antineoplastic agent exhibiting multiple action mechanisms. Recently, we found that after cytochrome P450 (CYP)-mediated oxidation ellipticine forms covalent DNA adducts. Ellipticine oxidation by isolated CYP and its binding to DNA is the target of this study.

Study of cytokinin metabolism using HPLC with radioisotope detection.

High performance liquid chromatography (HPLC) with an on-line flow-through radioactivity detector was used to monitor the metabolism of cytokinins ([3H]6-benzylaminopurine and [3H]6-benzylaminopurine riboside) after their incorporation into wheat seedlings. The production and conversion of individual metabolites was assayed within a short time interval (0.5-3 h).

Capillary electrophoresis of cytokinins and cytokinin ribosides.

A CE separation of cytokinins and cytokinin ribosides and some other purine and pyrimidine bases has been developed. Two electrolyte systems have been tested: 150 mM phosphoric acid, pH 1.8 and 50 mM sodium dodecylsulphate + 20 mM borate, pH 9.

Partial auxin deprivation affects endogenous cytokinins in an auxin-dependent, cytokinin-independent tobacco cell strain.

The dynamics of individual endogenous cytokinins within the growth cycle (subculture interval) of an auxin-dependent and cytokinin-independent cell suspension culture ofNicotiana tabacum L. (strain VBI-0) were determined using high performance liquid chromatography and radioimmunoassay. In cells grown at an optimum auxin concentration the transient maxima of N(6)-(Δ(2)-isopentenyl)adenine and N(6)-(Δ(2)-isopentenyl)-adenosine correlated with the onset of cell division.

Yersinia enterocolitica in water and foods.

Yersinia enterocolitica (Ye) isolates from water (367 strains) and different foods (114 strains) were typed serologically and biochemically. In some samples, especially of water, several Ye serotypes were found simultaneously. A noteworthy finding was Ye 03, biotype 4 in tinned meat paste that had already been on sale without, however, any case of diarrhoeal disease being reported in the particular district.

A finding of Yersinia pseudotuberculosis in well water.

From a sample of non-treated, non chlorinated well water a Yersinia pseudotuberculosis II strain was isolated. The water had been apparently massively contaminated because the strain was isolated from 1 ml of the sample. The serotype II of Y.