Comparison between orientational and conformational orders in fluid lipid bilayers.

The orientational order as determined by 2H NMR and the infrared frequencies of the C--H stretching modes of the methylene groups have been measured for several systems (POPC, POPC/cholesterol and POPE), all in the fluid phase, and then were compared; this work reveals an unexpected linear correlation between them. This experimental result shows that both measurements are essentially sensitive to a common motion, most likely trans/gauche isomerisation. This new correlation with those already found in the literature suggest that several measurements related to the hydrophobic core of the fluid bilayer describe different aspects of a universal behavior. The correlation presented here does not extend to the lipid in gel phase where slower motions affect the NMR lineshape.

Chain length-dependent Pb(II)-coordination in phytochelatins.

UV/visible and circular dichroism (CD) spectroscopy have been used to study the binding of Pb(II) to plant metal-sequestering peptides, phytochelatins (PCs), with the structure (gamma Glu-Cys)2Gly, (gamma Glu-Cys)3Gly and (gamma Glu-Cys)4Gly. Saturation of the Pb(II)-induced charge-transfer bands indicated that both (gamma Glu-Cys)2Gly and (gamma Glu-Cys)3Gly bound one metal ion per peptide molecule. However, (gamma Glu-Cys)4Gly formed two distinct species with stoichiometries of one and two Pb(II) ions per peptide molecule, respectively. The optical spectra of Pb(II)1-(gamma Glu-Cys)4Gly were similar to those of Pb(II)1-(gamma Glu-Cys)3Gly, whereas the spectra of Pb(II)2-(gamma Glu-Cys)4Gly were similar to those of Pb(II)1-(gamma Glu-Cys)2Gly. Since cysteinyl thiolates are the likely ligands for Pb(II) in PCs, Pb(II) appears to form two-, three- and four-coordinate complexes with PCs depending on their chain length. Furthermore, Pb(II) may exhibit multiple coordination in longer chain PCs as indicated by the formation of two Pb(II)-binding species of (gamma Glu-Cys)4Gly. The transfer of Pb(II) from glutathione to PCs and from shorter chain to longer chain PCs is also demonstrated.

Optical spectroscopic and reverse-phase HPLC analyses of Hg(II) binding to phytochelatins.

Optical spectroscopy and reverse-phase HPLC were used to investigate the binding of Hg(II) to plant metal-binding peptides (phytochelatins) with the structure (gammaGlu-Cys)2Gly, (gammaGlu-Cys)3Gly and (gammaGlu-Cys)4Gly. Glutathione-mediated transfer of Hg(II) into phytochelatins and the transfer of the metal ion from one phytochelatin to another was also studied using reverse-phase HPLC. The saturation of Hg(II)-induced bands in the UV/visible and CD spectra of (gammaGlu-Cys)2Gly suggested the formation of a single Hg(II)-binding species of this peptide with a stoichiometry of one metal ion per peptide molecule. The separation of apo-(gammaGlu-Cys)2Gly from its Hg(II) derivative on a C18 reverse-phase column also indicated the same metal-binding stoichiometry. The UV/visible spectra of both (gammaGlu-Cys)3Gly and (gammaGlu-Cys)4Gly at pH 7.4 showed distinct shoulders in the ligand-to-metal charge-transfer region at 280-290 mm. Two distinct Hg(II)-binding species, occurring at metal-binding stoichiometries of around 1.25 and 2.0 Hg(II) ions per peptide molecule, were observed for (gammaGlu-Cys)3Gly. These species exhibited specific spectral features in the charge-transfer region and were separable by HPLC. Similarly, two main Hg(II)-binding species of (gammaGlu-Cys)4Gly were observed by UV/visible and CD spectroscopy at metal-binding stoichiometries of around 1.25 and 2.5 respectively. Only a single peak of Hg(II)-(gammaGlu-Cys)4Gly complexes was resolved under the conditions used for HPLC. The overall Hg(II)-binding stoichiometries of phytochelatins were similar at pH 2.0 and at pH 7.4, indicating that pH did not influence the final Hg(II)-binding capacity of these peptides. The reverse-phase HPLC assays indicated a rapid transfer of Hg(II) from glutathione to phytochelatins. These assays also demonstrated a facile transfer of the metal ion from shorter- to longer-chain phytochelatins. The strength of Hg(II) binding to glutathione and phytochelatins followed the order: gammaGlu-Cys-Gly<(gammaGlu-Cys)2Gly<(gammaGlu-Cy s)3Gly<(gamma Glu-Cys)4Gly.