Nikola Paul Chmel

Self-assembling optically pure Fe(A-B)3 chelates.

Optically pure, single diastereomer fac-tris(diimine) complexes of Fe(II) are available from a remarkably facile one-pot procedure using a range of readily available (R)-2-phenylglycinol derivatives.

Origins of stereoselectivity in optically pure phenylethaniminopyridinetris-chelates M(NN′)3n+ (M = Mn, Fe, Co, Ni and Zn)

One-pot reactions of 2-pyridinecarboxaldehyde, chiral phenylethanamines and Fe(II) give single diastereomer fac diimine complexes at thermodynamic equilibrium so that no chiral separations are required (d.r. > 200 : 1). The origins of this stereoselectivity are partly steric and partly a result of the presence of three sets of inter-ligand parallel-offset π-stacking interactions. Mn(II), Co(II), Co(III), Ni(II) and Zn(II) give similar fac structures, alongside the imidazole analogues for Fe(II). While most of the complexes are paramagnetic, the series of molecular structures allows us to assess the influence of the π-stacking present, and there is a strong correlation between this and the M-N bond length. Fe(II) is close to optimal. For the larger Zn(II) ion, very weak π-stacking leads to poorer measured stereoselectivity (NMR) but this is improved with increased solvent polarity. The mechanism of stereoselection is further investigated via DFT calculations, chiroptical spectroscopy and the use of synthetic probes.

Membrane protein extraction and purification using styrene-maleic acid (SMA) co-polymer: effect of variations in polymer structure

The use of styrene-maleic acid (SMA) copolymers to extract and purify transmembrane proteins, while retaining their native bilayer environment, overcomes many of the disadvantages associated with conventional detergent-based procedures. This approach has huge potential for the future of membrane protein structural and functional studies. In this investigation, we have systematically tested a range of commercially available SMA polymers, varying in both the ratio of styrene and maleic acid and in total size, for the ability to extract, purify and stabilise transmembrane proteins. Three different membrane proteins (BmrA, LeuT and ZipA), which vary in size and shape, were used. Our results show that several polymers, can be used to extract membrane proteins, comparably to conventional detergents. A styrene:maleic acid ratio of either 2:1 or 3:1, combined with a relatively small average molecular mass (7.5-10 kDa), is optimal for membrane extraction, and this appears to be independent of the protein size, shape or expression system. A subset of polymers were taken forward for purification, functional and stability tests. Following a one-step affinity purification, SMA 2000 was found to be the best choice for yield, purity and function. However, the other polymers offer subtle differences in size and sensitivity to divalent cations that may be useful for a variety of downstream applications.

Organic-soluble optically pure anionic metal complexes PPh4[MIII(S,S-EDDS)]·2H2O (M = Fe, Co, Cr)

The first organic-soluble, optically and diastereomerically pure EDDS metal complexes have been synthesised. A number of synthetic approaches were attempted, but finally the tetraphenylphosphonium series emerged as providing readily accessible compounds of trivalent Cr, Fe and Co in reasonable yields via the silver salts without the need to perform ion-exchange chromatography. The species PPh(4)[M(III)(S,S-EDDS)] are very soluble in methanol, acetonitrile and even THF but isolation was facilitated by addition of stoichiometric water giving the highly crystalline but still conveniently soluble title compounds. The structures of the three isomorphous crystals comprise H(2)O-bridged extended hydrogen bonded structures with large channels occupied by the counterion molecules. The magnetic properties and circular dichroism spectra are reported along with comparative data for water-soluble NH(4)[Fe(III)(S,S-EDDS)]. Phase purity (and hence diastereomeric purity) in the paramagnetic systems is assessed through powder XRD. The practical utility of this type of compound was confirmed by optical resolution of (+/-)-[Ru(II)(bpy)(3)]Cl(2).

Redox-active and DNA-binding coordination complexes of clotrimazole.

DNA interactions of anticancer mononuclear Cu(2+), Co(2+), Zn(2+), and Ni(2+) complexes with the biologically active ligand clotrimazole (clotri) are reported. To fully characterize DNA binding modes for these complexes of the formulae [M(clotri)2Cl2]·nH2O (1-4), [M(clotri)2Br2]·nH2O (5,6), [M(clotri)3NO3]NO3·nH2O (9), and [M(clotri)3(NO3)2] (10), circular dichroism (CD) and linear dichroism (LD) spectroscopy, UV melting experiments, atomic force microscopy (AFM) and ethidium bromide (EtBr) displacement methods were used. Results indicate mixed electrostatic interactions, possibly through groove binding, that result in accretion and coiling of DNA. Electrochemical studies indicate that the Cu(2+) complex 9 readily reduces to the reactive-oxygen-species-generating Cu(+), which oxidatively damages DNA. There is a subtle correlation between log P values, calculated electrostatic potentials, and cytotoxicity of the complexes. The extent of cell-nucleus DNA-metal adduct formation in the HeLa cervix-uterine carcinoma cell line does not necessarily correlate with cytotoxicity, indicating that the nature of DNA lesions may be crucial to activity.

Jahn–Teller effects on π-stacking and stereoselectivity in the phenylethaniminopyridine tris-chelates Cu(NN′)32+

Optically pure phenylethaniminopyridine (S(C)-L) tris-chelates of Fe(II) and other first row transition metal systems have previously been shown to give exclusively the fac structures in the solid state. Here it is shown by powder X-ray diffraction that the complex [CuL(3)][ClO(4)](2) crystallises exclusively as the mer isomer, although--for a given absolute configuration of the ligand--of the same helicity (Δ/Λ) as that displayed by the other metal complexes. The similar ligand R(C)-L(F), which contains a peripheral (19)F spin label, gave [CuL(F)(3)][ClO(4)](2) which also adopts exclusively the mer structure in the crystal, but is shown by NMR spectroscopy to have a fac:mer ratio of 1:6 in solution at low temperature. Molecular mechanics calculations for a number of isomers and conformers are consistent with the presence of such a mixture of isomers in solution for both complexes. The origin of the difference in behaviour between Fe(II) and Cu(II) is the presence of a Jahn-Teller distortion (and the generally longer M-N bonds) in the Cu(II) complexes. This disturbs intra-ligand π-stacking, leading to the poor fac/mer stereoselectivity while leaving enantioselectivity Δ/Λ apparently unaffected.

Circular and Linear Dichroism Spectroscopy for the Study of Protein–Ligand Interactions

Circular dichroism (CD) is the difference in absorption of left and right circularly polarized light, usually by a solution containing the molecules of interest. A non-zero signal for solutions is only measured for chiral molecules such as proteins whose mirror image is not superposable on the original molecule. A CD spectrum provides information about the bonds and structures responsible for the chirality. When a small molecule (or ligand) binds to a protein, it acquires an induced CD (ICD) spectrum through chiral perturbation to its structure or electron rearrangements (transitions). The wavelengths of this ICD are determined by the ligands own absorption spectrum, and the intensity of the ICD spectrum is determined by the strength and geometry of its interaction with the protein. Thus, ICD can be used to probe the binding of ligands to proteins. This chapter contains an outline of how to perform protein CD and ICD experiments, together with some of the issues relating to experimental design and implementation. Addition of a quarter wave plate to a CD spectropolarimeter converts it to a linear dichroism (LD) spectrometer. When protein samples are aligned either in flow (as for fibers or membrane proteins in liposomes) or on surfaces the orientations of ligands with respect to the protein backbone or other subunits can be determined.

Chiral Semiconductor Phases: The Optically Pure D3[MIII(S,S-EDDS)]2 (D = TTF, TSF) Family

A new family of optically pure tetrathiafulvalenium and tetraselenafulvalenium salts, D(3)[M(III)(S,S-EDDS)](2)·nH(2)O (where D = TTF, TSF; M = Co, Fe, Cr; EDDS = ethylenediaminedisuccinato), were synthesized electrochemically. These phases are semiconductors with conductivities between 6.9 × 10(-6) and 1.3 × 10(-5) S·cm(-1) (E(a)ca. 0.3 eV) for TTF and 2.8 × 10(-4) to 2.8 × 10(-5) S·cm(-1) (E(a)ca. 0.1 eV) for TSF compounds. While some crystals suffer from twinning, other well resolved structures consist of TTF/TSF stacks which, under the influence of the chiral anion, exhibit a periodic undulation described by an elliptical helix. The crystallographic data, along with computational work, indicate charge localization in the semiconducting motifs.

Fluorescence detected linear dichroism spectroscopy: A selective and sensitive probe for fluorophores in flow-oriented systems

Fluorescence detection typically enhances sensitivity and selectivity for fluorescent analytes. The potential for combining fluorescence detection with flow orientation of the sample in the normal configuration of linear dichroism experiments is explored in this work by measuring the fluorescence emitted from flow-orientated DNA-bound ligands and M13 bacteriophage. Data for ethidium bromide, Hoechst 33258, and 4,6-diamidino-2-phenyindole are presented. The theoretical basis of the technique is also presented for instruments running in both the fixed direct-current mode, which is the normal operation mode of circular dichroism spectropolarimeters, and also in fixed high-tension voltage mode. The role of the stray light reaching the detector that results in a spectral shape in fixed direct current mode that resembles the shape of a linear dichroism spectrum, rather than the expected reduced linear dichroism, is also explored.

Considerations of noise and measurement reproducibility of circular dichroism measurements using Na[CoIII(EDDS)].

Although circular dichroism (CD) spectroscopy has become ubiquitous in biological, chemical, and pharmacological laboratories, several issues with instrument calibration and standardization, measurements quality, and reproducibility still remain. In the context of validating and developing a wide-wavelength-range stable standard for CD that is available in both enantiomeric forms, (R,R)- and (S,S)-Na[Co(III)(EDDS)] (where EDDS is ethylenediaminedisuccinato), we have become aware of sources of error in CD spectroscopy in addition to instrument calibration and user issues. This article summarizes those errors and suggests procedures for minimizing them by averaging over sufficient independent data accumulations with appropriate parameters.