Chiral Ligand-Exchange Resolution of Underivatized Amino Acids on a Dynamically Modified Stationary Phase for RP-HPTLC

The synthesis of Spi(tau-dec), derived from the selective alkylation of L-spinacine (4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-6-carboxylic acid) at the tau-nitrogen of its heteroaromatic ring, with a linear hydrocarbon chain of 10 carbon atoms, is described here for the first time. Spi(tau-dec) was successfully employed in the past to prepare home-made chiral columns for chiral ligand-exchange high-performance liquid chromatography. In the present article a new method is described, using Spi(tau-dec) as a chiral selector in high-performance thin-layer chromatography (HPTLC): commercial hydrophobic plates were first coated with Spi(tau-dec) and then treated with copper sulfate. The performance of this new chiral stationary phase was tested against racemic mixtures of aromatic amino acids, after appropriate optimization of both the conditions of preparation of the plates and the mobile phase composition. The enantioselectivity values obtained for the studied compounds were higher than those reported in the literature for similar systems. The method employed here for the preparation of chiral HPTLC plates proved practical, efficient, and inexpensive

Nickel complexes of dipeptides: a thermodynamic and spectroscopic study

Complex formation equilibria of some dipeptides (glycyl-alanine, glycyl-lysine, alanyl-alanine, alanyl-lysine, histidyl-alanine and histidyl-lysine) with the Ni(II) ion have been studied in aqueous solution at I=0.1 mol dm−3 (KNO3) and T=25°C. Protonation and complex formation constants have been potentiometrically determined. The structure of the main complex species is discussed on the basis of thermodynamic data obtained by direct calorimetry, CD and absorption spectra. The participation of the amide nitrogen in complex formation is suggested, while no evidence supports the participation of the NH2 side group of lysine in coordination

Quantitative SPLITT fractionation of lagoon sediments

In this work, SPLITT Fractionation (split flow thin cell) is used to sort hydrodynamically sedimented particles coming from the Sacca di Goro, a lagoon-like system close to the Po River delta (Italy). First the possibility of performing quantitative mass separations with a SPLITT cell apparatus was checked on a standard silica sample of known particle size distribution (PSD). Environmental sediment samples and relative SPLITT sub-fractions were subject to Inductive Coupled Plasma – Atomic Emission Spectroscopy (ICP-AES) characterization for the following elements: Al, Fe, Cd, Cr, Cu, Ni, Mn, Pb and Zn. The distribution of these metals by particle size fractions has been investigated. The accuracy of the entire separation procedure has been also evaluated

Formation equilibria of nickel complexes with glycyl-histidyl-lysine and two synthetic analogues

Complex-formation equilibria between the Ni(II) ion and the natural tripeptide glycyl-L-histidyl-L-lysine have been investigated. Two synthetic analogues, where the histidine residue has been substituted with L-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-6-carboxylic acid (L-Spinacine) and L-1,2,3,4-tetrahydro-isoquinolin-3-carboxylic acid (Tic), respectively, have been considered, as well. Different experimental techniques have been employed: potentiometry, calorimetry, visible spectrophotometry and CD spectroscopy. Structural hypotheses on the main complex species are suggested. Evidences on the formation of tetrameric species with the first ligand are shown. No involvement of the side-chain amino group of lysine residue in metal ion coordination was found

THE UNUSUAL STABILIZATION OF THE Ni2+ AND Cu2+ COMPLEXES WITH NSFRY

The binding mode provided by an unprotected peptide with non-coordinating side-chains is simple and well understood. However, when particular residues are inserted into the peptide sequence, they can have a significant impact on the stability of the formed complexes. The presence of non-bonding side chains of amino acids close to the metal binding centre in the peptide/protein can provide special interactions which result in increasing the stabilization of the formed species. Moreover, these interactions can play a crucial role in generating particular protein structures and in influencing biological activity. In the present paper it is shown how peptides with no specific predisposition for metal binding, like ANF peptides, can form metal complexes with a very high thermodynamic stability. For better understanding this peculiar behavior, a combined pH-metric and spectroscopic method was used to determine the stability and the solution structure of Cu2+ and Ni2+ complexes with NSFRY-NH2 (ANF peptide) and a series of analogue peptides. All obtained data support the hypothesis that the complex-formation process is very similar for both metal ions and all the ligands, involving some intramolecular interactions among the different side chains. The two-dimensional NMR analysis of nickel complexes showed the occurrence of many inter-residue correlations and suggested the presence a direct interaction between the d electrons of the metal ion and the π-ring system of the aromatic side-chains of the ligand

Cu(II) ion coordination to the pentadecapeptide model of SPARC copper-binding site

SPARC (Secreted Protein, Acidic and Rich in Cysteine) is a matricellular glycoprotein with many biological functions: it mediates the interactions between cells and the extracellular matrix, playing a role in angiogenesis, tumorigenesis, caractogenesis and wound healing. Proteolysis of SPARC gives rise to a number of oligopeptides which can regulate angiogenesis in vivo and the biological activity of which has been related to their association with endogenous or exogenous copper ion. Human SPARC consists of three distinct modules. Module II is follistatinlike and contains two copper binding sites, the strongest of which—the cationic region 2 (amino acids 114–130)— contains the sequence Gly–His–Lys. In order to shed more light on the biological role of metal complexes formed by SPARC and its fragments, more information is needed on their stoichiometry, stability and structure in solution. In the present paper a potentiometric and spectroscopic investigation on Cu(II) complexes with the SPARC114–128 fragment, protected at both its amino and carboxylic ends, is reported. This peptide (Ac–TLEGTKKGHKLHLDY– NH2) constitutes a good model to the strong copper-binding site of the protein. The whole experimental data suggest that complex-formation is started by the two His residues, subsequently involving up to three amido nitrogens, as pH increases. The coordination of the two histydyl imidazoles is able to promote amide ionisation in the physiological pH range and this could be the key to the SPARC affinity for Cu(II) ion