Anastasios Troganis

Do strong intramolecular hydrogen bonds persist in aqueous solution? Variable temperature gradient 1H, 1H–13C GE-HSQC and GE-HMBC NMR studies of flavonols and flavones in organic and aqueous mixtures

Abstract Intramolecular hydrogen bonds in crystals and in apolar media are well documented, however, the degree to which they persist in aqueous solution is controversial. We report here variable temperature gradient 1 H, 1 H– 13 C Gradient Enhanced Heteronuclear Single Quantum Correlation (GE-HSQC) and Gradient Enhanced Heteronuclear Multiple Bond Coherence (GE-HMBC) NMR studies of the flavonols quercetin and kaempferol and the flavone luteolin, in organic solvents and in mixtures of organic–aqueous solutions. It is demonstrated that the strong intramolecular hydrogen bond of the –CO(4) and –OH(5) moieties persists over a wide range of aqueous mixtures and, thus, provide a rare example of non-charged intramolecular hydrogen bonds, which is not overwhelmed by solvation with protic solvents, in particular in aqueous solution.

Treatment of experimental allergic encephalomyelitis (EAE) by a rationally designed cyclic analogue of myelin basic protein (MBP) epitope 72-85.

In this report the rational design, synthesis and pharmacological properties of an amide-linked cyclic antagonist analogue of the guinea pig myelin basic protein epitope MBP(72-85) are described. Design of the potent cyclic analogue was based on 2D NOESY nuclear magnetic resonance and molecular dynamics studies carried out in the linear antagonist Ala81MBP(72-85). The cyclic antagonist completely prevented the induction of experimental allergic/autoimmune encephalomyelitis when coinjected with linear and cyclic agonist analogues MBP(72-85) and cyclo(2-9)MBP(72-85).

Synthetic analogs for oxovanadium(IV/V)-glutathione interaction: an NMR, EPR, synthetic and structural study of oxovanadium(IV/V) compounds with sulfhydryl-containing pseudopeptides and dipeptides.

The reaction of [VO(CH3COO)2(phen)] (phen = 1,10-phenanthroline) with the sulfhydryl-containing pseudopeptides (scp), N-(2-mercaptopropionyl)glycine (H3mpg), N-(2-mercaptopropionyl)cysteine (H4m2pc), N-(3-mercaptopropionyl)cysteine (H4m3pc) and the dipeptides glycylglycine (H2glygly) and glycyl-L-alanine (H2glyala), in the presence of triethylamine, results in the formation of the compounds Et3NH[VO(mpg)(phen)] (1), (Et3NH)2[VO(m2pc)] (4), [(Et3NH)2[VO(m3pc) (5), [VO(glygly)(phen)] x 2CH3OH (2 x 2CH3OH) and [VO(glyala)(phen)] x CH3OH (3 x CH3OH). Evidence for the molecular connectivity in 2 x CH3OH was established by X-ray crystallography, showing the vanadium(IV) atom ligated to a tridentate glygly2- ligand at the N(amine), N(peptide) and O(carboxylato) atoms. Combination of the correlation plot of the EPR parameters gz versus Az, together with the additivity relationship supported the prediction of the equatorial donor atom sets of the V(IV)O2+ center at various pH values for the V(IV)O2+-glutathione system considered in this study. Model NMR studies (interaction of vanadium(V) with the scp H3mpg) showed that there is a possibility of vanadium(V) ligation to glutathione.

Towards a consensus structure of hypericin in solution: direct evidence for a single tautomer and different ionization states in protic and nonprotic solvents by the use of variable temperature gradient 1H NMR

Abstract Hypericin, a meso -naphthodianthrone derivative, displays two types of electronic spectra in organic solvents, attributed to the existence of two tautomeric structures. Variable temperature gradient 1 H NMR studies demonstrate the occurrence of only one 7,14-dioxo tautomeric form, for the molecule of hypericin in protic and in nonprotic solvents, differing only in the degree of ionization of the 4-hydroxyl group in the bay region.

Thermodynamic origin ofcis/trans isomers of a proline-containing β-turn model dipeptide in aqueous solution: A combined variable temperature1H-NMR, two-dimensional1H,1H gradient enhanced nuclear overhauser effect spectroscopy (NOESY), one-dimensional steady-state intermolecular13C,1H NOE, and molecular dynamics study

The cis/trans conformational equilibrium of the two Ac-Pro isomers of the beta-turn model dipeptide [13C]-Ac-L-Pro-D-Ala-NHMe, 98% 13C enriched at the acetyl carbonyl atom, was investigated by the use of variable temperature gradient enhanced 1H-nmr, two-dimensional (2D) 1H,1H nuclear Overhauser effect spectroscopy (NOESY), 13C,1H one-dimensional steady-state intermolecular NOE, and molecular dynamics calculations. The temperature dependence of the cis/trans Ala(NH) protons are in the region expected for random-coil peptides in H2O (delta delta/delta T = -9.0 and -8.9 ppb for the cis and trans isomers, respectively). The trans NH(CH3) proton indicates smaller temperature dependence (delta delta/delta T approximately -4.8 ppb) than that of the cis isomer (-7.5 ppb). 2D 1H,1H NOESY experiments at 273 K demonstrate significant NOEs between ProH alpha-AlaNH and AlaNH-NH(R) for the trans isomer. The experimental NOE data, coupled with computational analysis, can be interpreted by assuming that the trans isomer most likely adopts an ensemble of folded conformations. The C-CONH(CH3) fragment exhibits significant conformational flexibility; however, a low-energy conformer resembles closely the beta II-turn folded conformations of the x-ray structure of the related model peptide trans-BuCO-L-Pro-Me-D-Ala-NHMe. On the contrary, the cis isomer adopts open conformations. Steady-state intermolecular solute-solvent (H2O) 13C,1H NOE indicates that the water accessibility of the acetyl carbonyl carbons is nearly the same for both isomers. This is consistent with rapid fluctuations of the conformational ensemble and the absence of a highly shielded acetyl oxygen from the bulk solvent. Variable temperature 1H-nmr studies of the cis/trans conformational equilibrium indicate that the trans form is enthalpically favored (delta H degree = -5.14 kJ mole-1) and entropically (delta S degree = -5.47 J.K-1.mole-1) disfavored relative to the cis form. This demonstrates that, in the absence of strongly stabilizing sequence-specific interresidue interactions involving side chains and/or charged terminal groups, the thermodynamic difference of the cis/trans isomers is due to the combined effect of intramolecular and intermolecular (hydration) induced conformational changes.

Novel lipophilic amidate oxorhenium and oxotechnetium complexes as potential brain agents: synthesis, characterization and biological evaluation

Abstract. Novel oxorhenium and oxotechnetium complexes based on the tetradentate 1-(2-hydroxybenzamido)-2-(pyridinecarboxamido)benzene, H3L, ligand have been synthesized and characterized herein. Thus, by reacting equimolar quantities of the triply deprotonated ligand L3– with the suitable MO3+ precursor, the following neutral MOL complexes could be easily produced following similar synthetic routes: M=Re (1), M=99gTc (2), and M=99mTc (3). Complexes 1 and 2, prepared in macroscopic amounts, were chemically characterized and their structure determined by single-crystal X-ray analysis. They are isostructural metal chelates, adopting a distorted square pyramidal geometry around the metal. The N3O donor atom set of the tetradentate ligand defines the basal plane and the oxygen atom of the M=O core occupies the apex of the pyramid. Complex 3 forms quantitatively at tracer level by mixing the H3L ligand with Na99mTcO4 generator eluate in aqueous alkaline media and using tin chloride as reductant in the presence of citrate. Its structure was established by chromatographic comparison with prototypic complexes 1 and 2 using high-performance liquid chromatographic techniques. When challenged with excess glutathione in vitro, complex 3 is rapidly converted to hydrophilic unidentified metal species. Tissue distribution data after administration of complex 3 in vivo revealed a significant uptake and retention of this compound in brain tissue.

Involvement of phospholipids in apolipoprotein B modification during low density lipoprotein oxidation

An increased amount of phospholipids remained attached on delipidated apolipoprotein B originated from oxidized low density lipoprotein (LDL). 31P nuclear magnetic resonance analysis of such apolipoprotein showed an organic phosphorus peak at −0.55 ppm, which suggests the formation of adducts (most probably Schiff bases) of oxidized phospholipids with apolipoprotein B. The above reaction occurs in parallel with the hydrolysis of oxidized phospholipids, catalyzed by the LDL-attached platelet-activating factor acetylhydrolase, and may contribute to the proatherogenic effect of oxidatively modified low density lipoprotein.

Arg side-chain-backbone interactions evidenced in model peptides by 17O-NMR spectroscopy.

The guanidinium group of arginine possesses a variety of biochemical functions, either by participating in direct interactions in recognition processes, or by stabilizing secondary structures. Three model compounds, selectively (17)O enriched, Ac-Arg-Ala-[(17)O]Pro-NH(2) (1), Piv-Arg-Pro-[(17)O]Gly-NH(2) (2) (C-terminal segment of the luteinizing hormone releasing hormone), and Piv-Nle-Pro-[(17)O]Gly-NH(2) (3), were prepared and studied by (17)O-nmr spectroscopy. A direct hydrogen-bonded interaction between the Arg side chain and the carbonyl main chain carboxy-terminus was found, thus confirming the tendency of Arg to participate in proton-acceptor functions.

Two of a kind but different: Luminescent carbon quantum dots from Citrus peels for iron and tartrazine sensing and cell imaging

Citrus sinensis and Citrus limon peels were used to synthesize two different kinds of carbon quantum dots (CQDs) via an unsophisticated and inexpensive carbonization procedure. The proposed synthesis is straightforward and adheres to the principles of green chemistry since no organic solvents are used and no toxic by-products are formed, while the residual resources employed facilitate the large scale synthesis of dots. The Citrus sinensis and Citrus limon peels are proved to be excellent precursors for the synthesis of CQDs with highly practical applications. The CQDs display strong excitation-independent, blue fluorescence, which is stable over time. Splendid water dispersibility, photostability and stability over a wide range of pH are some of the main advantages of the CQDs, which enable them to be used as a fluorescent probes. Although many of their features are alike, our findings demonstrate that each kind of the CQDs lend itself to quite distinct analytical applications. The developed fluorescent probes possess high potential for sensitive and selective detection of Fe3+ (Citrus sinensis CQDs) and tartrazine (Citrus limon CQDs) via a quenching mechanism. The decrease in fluorescence intensity is in linear relationship with the concentrations of Fe3+ and tartrazine in the ranges of 0.01-1.0μM and 0.6-23.5μΜ, respectively. Moreover, their low cytotoxicity reinforces their applicability towards cell bioimaging and intracellular detection of Fe3+, which were further studied.

Multinuclear (13C, 17O, 57Fe) NMR studies of carbonmonoxy heme proteins and synthetic model compounds

13C, 17O and 57Fe NMR spectra of several carbonmonoxy hemoprotein models with varying polar and steric effects of the distal organic superstructure, constraints of the proximal side, and porphyrin ruffling are reported. Both heme models and heme proteins obey a similar excellent linear delta(13C) versus nu(C-O) relationship which is primarily due to modulation of pi-back-bonding from the Fe d(pi) to CO pi* orbital by the distal pocket polar interactions. The lack of correlation between delta(13C) and delta(17O) suggests that the two probes do not reflect a similar type of electronic and structural perturbation. delta(17O) is not primarily influenced by the local distal field interactions and does not correlate with any single structural property of the Fe-C-O unit; however, atropisomerism and deformation of the porphyrin geometry appear to play a significant role. 57Fe shieldings vary by nearly 900 ppm among various hemes and an excellent correlation was found between delta(57Fe) and the absolute crystallographic average displacement of the meso carbon atoms, /Cm/, relative to the porphyrin core mean plane. The excellent correlation between iron-57 shieldings and the average shieldings of the meso carbons of the porphyrin skeleton of TPP derivatives suggests that the two probes reflect a similar type of electronic and structural perturbation which is primarily porphyrin ruffling.