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The metal ion coordination abilities of reduced and oxidized glutathione are reviewed. Reduced glutathione (GSH) is a very versatile ligand, forming stable complexes with both hard and soft metal ions. Several general binding modes of GSH are described. Soft metal ions coordinate exclusively or primarily through thiol sulfur. Hard ones prefer the amino acid-like moiety of the glutamic acid residue. Several transition metal ions can additionally coordinate to the peptide nitrogen of the γ-Glu-Cys bond. Oxidized glutathione lacks the thiol function. Nevertheless, it proves to be a surprisingly efficient ligand for a range of metal ions, coordinating them primarily through the donors of the glutamic acid residue.
The review presents specific interactions that occur in complexes of Cu(ll) ions with peptides composed only of amino acids with nonco-ordinating side chains. Three classes of such peptides are discussed. The first type (NSFRY analogues) is characterised by the presence of a specific combination of bulky and aromatic residues, leading to a formation of multiple weak interactions around Cu(II) that result in an extremely high stability of complexes. The second class is composed of complexes of vasopressins and oxytocins, achieving superstability through a pre-conformation in the peptide molecule. The third group are oligopeptides containing one or two proline residues. These peptides form exotic macrochelate loops with Cu(ll) in a result of the break-point effect of Pro residues. Particular emphasis in the review was given to stability constants of complexes, compared to oligoglycine or oligoalanine peptides.
Asymmetrical diadenosine 5',5''-P 1P 4 tetraphosphate (Ap4A) hydrolases are key enzymes controlling the in vivo concentration of Ap4A - an important signaling molecule involved in regulation of DNA replication and repair, signaling in stress response and apoptosis. Sequence homologies indicate that the genome of the model plant Arabidopsis thaliana contains at least three open reading frames encoding presumptive Ap4A hydrolases: At1g30110, At3g10620, and At5g06340. In this work we present efficient overexpression and detailed biochemical characteristics of the AtNUDX25 protein encoded by the At1g30110 gene. Aided by the determination of the binding constants of Mn(Ap4A) and Mg(Ap4A) complexes using isothermal titration calorimetry (ITC) we show that AtNUDX25 preferentially hydrolyzes Ap4A in the form of a Mn 2+ complex.
Thimerosal (TH), an ethylmercury complex of thiosalicylic acid has been used as preservative in vaccines. Inspired by a known high affi nity of mercury for thiol groups, we examined whether the presence of L-cysteine (Cys), D,L-homocysteine (Hcy), Nacetyl cysteine (NAC), L-methionine (Met) and glutathione (GSH) in extracellular space could infl uence the viability, intracellular calcium concentration ([Ca2+]i ) and mitochondrial membrane potential in rat cerebellar granule cells. The cells were exposed to 500 nM TH for 48 h or 15 μM TH for 10 min. The loss of cells viability could be prevented partially or wholly, in a dose-dependent manner, by 60, 120 or 600 μM Cys, Hcy, NAC and GSH, but not by Met. The elevation in [Ca2+]i and mitochondrial potential induced by 25 μM TH were abolished by all compounds studied, except for Met, at 600 μM. The loss of the ethylmercury moiety from TH as a result of interaction with thiols studied was monitored by 1 Hand 199Hg-NMR spectroscopy. This extracelullar process may be responsible for the neuroprotection seen in cerebellar cell culture, but also provides a molecular pathway for redistribution of TH derived toxic ethylmercury in the organism.
In Escherichia coli, heterologous production of Schizosaccharomyces pombe phytochelatin synthase (PCS) along with overproduction of E. coli serine acetyltransferase (SAT) and gamma-glutamylcysteine synthase (gammaECS) was achieved and resulted in the accumulation of phytochelatins in bacterial cells. Overproduction of either gammaECS alone or simultaneous production of all three proteins in bacterial cells were accompanied by reduced growth rate in liquid cultures. Interestingly, bacteria overproducing either gammaECS or both SAT and gammaECS (with elevated level of gamma-glutamylcysteine but not of phytochelatins) were able to accumulate more cadmium per dry weight than the control. However, the most efficient cadmium accumulation was observed in bacteria with elevated levels of all three proteins: SAT, gammaECS and PCS. Therefore, “pushing” the entire pathway might be the most promising approach in modification of bacteria for potential bioremediation purposes because the level of intermediates, cysteine and glutathione, can limit the rate of production of phytochelatins. However, in such bacteria other metabolic process might become limiting for efficient growth.
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