Negative modulation of the GABAAρ1 receptor function by l-cysteine

l-Cysteine is an endogenous sulfur-containing amino acid with multiple and varied roles in the central nervous system, including neuroprotection and the maintenance of the redox balance. However, it was also suggested as an excitotoxic agent implicated in the pathogenesis of neurological disorders such as Parkinson′s and Alzheimer′s disease. l-Cysteine can modulate the activity of ionic channels, including voltage-gated calcium channels and glutamatergic NMDA receptors, whereas its effects on GABAergic neurotransmission had not been studied before. In the present work, we analyzed the effects of l-cysteine on responses mediated by homomeric GABAAρ1 receptors, which are known for mediating tonic γ-aminobutyric acid (GABA) responses in retinal neurons. GABAAρ1 receptors were expressed in Xenopus laevis oocytes and GABA-evoked chloride currents recorded by two-electrode voltage-clamp in the presence or absence of l-cysteine. l-Cysteine antagonized GABAAρ1 receptor-mediated responses; inhibition was dose-dependent, reversible, voltage independent, and susceptible to GABA concentration. Concentration-response curves for GABA were shifted to the right in the presence of l-cysteine without a substantial change in the maximal response. l-Cysteine inhibition was insensitive to chemical protection of the sulfhydryl groups of the ρ1 subunits by the irreversible alkylating agent N-ethyl maleimide. Our results suggest that redox modulation is not involved during l-cysteine actions and that l-cysteine might be acting as a competitive antagonist of the GABAAρ1 receptors. (Figure presented.).Fil: Beltrán González, Andrea Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Vicentini, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Calvo, Daniel Juan. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentin

Transamination of L-cysteine sulfinate in the growing rat.

The enzyme activities involved in the transamination of L-cysteine sulfinate (L-alanine 3-sulfinic acid), L-aspartate and L-cysteine were examined in fetal, neonatal and maternal rat liver and placenta. In fetal and neonatal rat liver, aminotransferase activity was most active with L-cysteine sulfinate as a substrate and was also active with L-aspartate, while activity with L-cysteine was very low. The activity of transamination of L-cysteine sulfinate in rat liver developed in parallel with that of L-aspartate and L-cysteine. The aminotransferase activity markedly increased after the 19th day of gestation, reaching the same value as adult liver on the 3rd day after birth. The ratios of transamination of L-cysteine sulfinate to that of L-aspartate and to that of L-cysteine were constant during development. These observations suggest that L-cysteine sulfinate, L-aspartate and L-cysteine are transaminated by the same enzyme in the rat liver during development. Since placental aminotransferase activity was extremely low compared with that of the liver, it was suggested that the placenta did not play an important role in the transamination of these amino acids during pregnancy.</p

Multi-vanadium substituted polyoxometalates as efficient electrocatalysts for the oxidation of l-cysteine at low potential on glassy carbon electrodes

The electrochemical behaviours of the sandwich-type complex [As2W18(VO)3O66]11- were studied in a pH 7 medium and compared with those of the three following Dawson-type vanadium-substituted complexes: [P2V2W16O62]8- (P2V2W16), [P2MoV2W15O62]8- (P2MoV2W15) and [P2V3W15O62]9- (P2V3W15). Electrochemistry shows that the sandwich-type POM contains 2 VIV centers and one VV center and must be formulated As2V2IVVW18, in agreement with titration, elemental analysis and magnetic measurements on this element.. All the POMs of this work proved efficient for the oxidation of L-cysteine. Comparison of the present results with those of mono-Vanadium substituted POMs indicates that accumulation of vanadium atoms in the POM framework is beneficial in the electrocatalytic process. In addition, the present work highlights the important influence of the POM structure in the electrocatalytic oxidation of L-cysteine. The remarkable outcome of this work is that the potential for the oxidation of L-cysteine in the presence of the selected POMs has been substantially driven in the negative direction compared to the case of glassy carbon alone, a feature which is associated with faster kinetics. The stability of the systems must also be pointed out

PCR random mutagenesis into Escherichia coli serine acetyltransferase: isolation of the mutant enzymes that cause overproduction of L-cysteine and L-cystine due to the desensitization to feedback inhibition

AbstractPCR random mutagenesis in the cysE gene encoding Escherichia coli serine acetyltransferase was employed to isolate the mutant enzymes that, due to a much less feedback inhibition by L-cysteine, cause overproduction of L-cysteine and L-cystine in the recombinant strains. The L-cysteine auxotrophic and non-utilizing E. coli strain was transformed with plasmids having the altered cysE genes. Then, several transformants overproducing L-cysteine were selected by detecting the halo formation of the L-cysteine auxotroph. The production test of amino acids and analysis of the catalytic property on the mutant enzymes suggest that the carboxy-terminal region of serine acetyltransferase plays an important role in the desensitization to feedback inhibition and the high level production of L-cysteine and L-cystine

Lifespan extension and increased resistance to environmental stressors by N-Acetyl-L-Cysteine in Caenorhabditis elegans

OBJECTIVE: This study was performed to determine the effect of N-acetyl-L-cysteine, a modified sulfur-containing amino acid that acts as a strong cellular antioxidant, on the response to environmental stressors and on aging in C. elegans. METHOD: The survival of worms under oxidative stress conditions induced by paraquat was evaluated with and without in vivo N-acetyl-L-cysteine treatment. The effect of N-acetyl-L-cysteine on the response to other environmental stressors, including heat stress and ultraviolet irradiation (UV), was also monitored. To investigate the effect on aging, we examined changes in lifespan, fertility, and expression of age-related biomarkers in C. elegans after N-acetyl-L-cysteine treatment. RESULTS: Dietary N-acetyl-L-cysteine supplementation significantly increased resistance to oxidative stress, heat stress, and UV irradiation in C. elegans. In addition, N-acetyl-L-cysteine supplementation significantly extended both the mean and maximum lifespan of C. elegans. The mean lifespan was extended by up to 30.5% with 5 mM N-acetyl-L-cysteine treatment, and the maximum lifespan was increased by 8 days. N-acetyl-L-cysteine supplementation also increased the total number of progeny produced and extended the gravid period of C. elegans. The green fluorescent protein reporter assay revealed that expression of the stress-responsive genes, sod-3 and hsp-16.2, increased significantly following N-acetyl-L-cysteine treatment. CONCLUSION: N-acetyl-L-cysteine supplementation confers a longevity phenotype in C. elegans, possibly through increased resistance to environmental stressors

Removal of acetaldehyde from saliva by mucoadhesive formulations containing cysteine and chlorhexidine diacetate: a possible approach to the prevention of oral cavity alcohol-related cancer

The aim of our work has been to develop buccoadhesive formulations (tablets) containing both L-cysteine and chlorhexidine diacetate and to verify their ability to reduce oral acetaldehyde produced after alcoholic drinks consumption

An XPS Study of the Ag-S interface of L-Cysteine Films on Silver Surface

L-cysteine has gained much attention as a versatile amino acid to create bioactive surfaces by assisting the bonding of proteins to metal surfaces. When L-cysteine interacts with metallic partners such as gold, silver and copper, the SH functional group is known to interact strongly with the metallic surface. The interaction of the L-cysteine with silver surfaces is particularly interesting because, L-cysteine adsorption on silver has been suggested to be stronger than on gold or copper surfaces and the interactions may strongly influence the formation of novel interface states of the L-cysteine-Ag interface. On the other hand, some studies report a weakening of silver-sulfur bond with increasing coverage. However, research has not been sufficiently addressed for experimental investigation to understand the interaction of L-cysteine with silver metallic surfaces. As the first step, we reported the results of experimental investigation of L-cysteine and silver interface electronic structure by thickness-dependent ultraviolet photoelectron spectroscopy (UPS) with a clear spectral feature in between Fermi edge and highest occupied molecular orbital (HOMO) of L-cysteine due to the formation of Ag-S bonding together with a weakening of the silver-sulfur bond with increasing of L-cysteine. In this study, the formation of Ag-S bond at the L-cysteine modified silver surface was systematically elucidated by X-ray photoelectron spectroscopy (XPS) for three different coverages, namely monolayer, two-layered and multi-layered. A prominent shoulder at 2475.2 eV of the main peak at 2473.2 eV was observed for the S 1s XPS spectrum for monolayer film, while only a single peak was observed in the case of two- layered and multi-layered. The spectral feature at 2475.2 eV can be attributed to the interaction of L-cysteine with silver. In addition, the disappearance of the spectral feature for the two-layered film can be attributed to the weakening of silver-sulfur bond by over layer of L-cysteine

Modification of the Size of Supported Clusters by Coadsorption of an Organic Compound: Gold and l-Cysteine on Rutile TiO(2)(110).

Using X-ray photoelectron spectroscopy we studied the coadsorption of the amino acid l-cysteine and gold on a rutile TiO(2)(110) surface under ultrahigh vacuum conditions. Irrespective of the deposition order, i.e., irrespective of whether l-cysteine or gold is deposited first, the primary interaction between l-cysteine and the gold clusters formed at the surface takes place through the deprotonated thiol group of the molecule. The deposition order, however, has a profound influence on the size of the gold clusters as well as their location on the surface. If l-cysteine is deposited first the clusters are smaller by a factor two to three compared to gold deposited onto the pristine TiO(2)(110) surface and then covered by l-cysteine. Further, in the former case the clusters cover the molecules and thus form the outermost layer of the sample. We also find that above a minimum gold cluster size the gold cluster/l-cysteine bond is stronger than the l-cysteine/surface bridging oxygen vacancy bond, which, in turn, is stronger than the gold cluster/vacancy bond

Anharmonic transitions in nearly dry L-cysteine I

Two special dynamical transitions of universal character have been recently observed in macromolecules at $T_{D}\sim 180 - 220$ K and $T^{*}\sim 100$ K. Despite their relevance, a complete understanding of the nature of these transitions and their consequences for the bio-activity of the macromolecule is still lacking. Our results and analysis concerning the temperature dependence of structural, vibrational and thermodynamical properties of the orthorhombic polymorph of the amino acid L-cysteine (at a hydration level of 3.5%) indicated that the two referred temperatures define the triggering of very simple and specific events that govern all the biochemical interactions of the biomolecule: activation of rigid rotors ($T<T^{*}$), phonon-phonon interactions with phonons of water dimer ($T^{*}<T<T_{D}$), and water rotational barriers surpassing ($T>T_{D}$).Comment: 4 pages, 4 figures, submitted to Physical Review Letter