Effect of substituents on complex stability aimed at designing new iron(III) and aluminum(III) chelators

The solution equilibria of iron(III) and aluminum(III) with two classes of hard ligands (catechol, salicylic acid and their nitro-derivatives) have been reliably studied by potentiometric, spectrophotometric and NMR spectroscopy. The effect of the nitro substituent on the binding properties of catechol and salicylic acid has been examined thoroughly. The inductive and resonance properties of the substituent that, as expected, lower the basicity of the phenolic and carboxylic groups, lead to a general decrease in both protonation and complex formation constants. This decrease causes an increase in pM of between 0.2 and 1.1 pM units for the nitro-substituted salicylates and of about 4 units for 4-nitrocatechol, with a significantly higher chelating efficacy. The influence of the substituent on catechol and salicylic acid is discussed in detail on the basis of conditional constants at pH 7.4.     

Structure and in vitro activities of a Copper II-chelating anionic peptide from the venom of the scorpion Tityus stigmurus

Anionic Peptides are molecules rich in aspartic acid (Asp) and/or glutamic acid (Glu) residues in the primary structure. This work presents, for the first time, structural characterization and biological activity assays of an anionic peptide from the venom of the scorpion Tityus stigmurus, named TanP. The three-dimensional structure of TanP was obtained by computational modeling and refined by molecular dynamic (MD) simulations. Furthermore, we have performed circular dichroism (CD) analysis to predict TanP secondary structure, and UV–vis spectroscopy to evaluate its chelating activity. CD indicated predominance of random coil conformation in aqueous medium, as well as changes in structure depending on pH and temperature. TanP has chelating activity on copper ions, which modified the peptide’s secondary structure. These results were corroborated by MD data. The molar ratio of binding (TanP:copper) depends on the concentration of peptide: at lower TanP concentration, the molar ratio was 1:5 (TanP:Cu²⁺), whereas in concentrated TanP solution, the molar ratio was 1:3 (TanP:Cu²⁺). TanP was not cytotoxic to non-neoplastic or cancer cell lines, and showed an ability to inhibit the in vitro release of nitric oxide by LPS-stimulated macrophages. Altogether, the results suggest TanP is a promising peptide for therapeutic application as a chelating agent.     

Effect of S-100 protein on assembly of brain microtubule proteins in vitro

S-100 protein inhibits the assembly of brain microtubule proteins in vitro in the presence of 10 microM free Ca2+. The S-100 effect is generally greater on the rate than on the extent of assembly, and even greater as the microtubule protein concentration decreases and the time of preincubation between S-100 and microtubule proteins before GTP addition increases, at a given S-100/tubulin dimer molar ratio. The S-100 effect is greatly enhanced in the presence of physiological concentrations of K+ and is completely reversed by EGTA.     

Effects of Cadmium in Stylonychia lemnae, Stylonychia notophora and Oxytricha granulifera: Isolation of a Cadmium-Binding Protein

ABSTRACT The effects of cadmium on three ciliates are reported here. Cultures of Stylonychia lemnae, Stylonychia notophora and Oxytricha granulifera were treated with different doses of Cd according to tolerance. The two species of Stylonychia are very sensitive to the metal, white O. granulifera tolerates higher doses. Adding 50 μM of Cd to the medium did not damage cells. The accumulated metal is almost totally present in the particulate fraction after day 3. Two Cd-Zn linking fractions were separated from the soluble fraction of culture treated on day 1. The first protein linking 17 μg Cd/mg showed an ultraviolet absorption spectrum similar to that of Cd-thioneins. Preliminary amino acid analyses indicated that it contained 13% cysteine. The second protein, linking 60 μg Cd/mg, was a glycoprotein. Its ultraviolet absorption spectrum and amino acid analysis showed that this binding protein was far from being a metallothionein: its cysteine content was very low and aromatic and cyclic residues were present. This Cd-linking compound seems to be unique, since it was very different both from metallothioneins and chelatins isolated by other protozoa. The protective role of these chelating proteins is discussed.