pH-dependence of the specific binding of Cu(II) and Zn(II) ions to the amyloid-β peptide

Metal ions like Cu(II) and Zn(II) are accumulated in Alzheimer's disease amyloid plaques. The amyloid-β (Aβ) peptide involved in the disease interacts with these metal ions at neutral pH via ligands provided by the N-terminal histidines and the N-terminus. The present study uses high-resolution NMR spectroscopy to monitor the residue-specific interactions of Cu(II) and Zn(II) with (15)N- and (13)C,(15)N-labeled Aβ(1-40) peptides at varying pH levels. At pH 7.4 both ions bind to the specific ligands, competing with one another. At pH 5.5 Cu(II) retains its specific histidine ligands, while Zn(II) seems to lack residue-specific interactions. The low pH mimics acidosis which is linked to inflammatory processes in vivo. The results suggest that the cell toxic effects of redox active Cu(II) binding to Aβ may be reversed by the protective activity of non-redox active Zn(II) binding to the same major binding site under non-acidic conditions. Under acidic conditions, the protective effect of Zn(II) may be decreased or changed, since Zn(II) is less able to compete with Cu(II) for the specific binding site on the Aβ peptide under these conditions.     

A Revision of the Chinese Athyrium Roth II—Enumeration of the Species (1)

About 300 “species” names of Athyrium from China were published. They are preliminarily treated as 117 species with a number of varieties and hybrids. The complete enumeration will be reported in four parts. The present paper is part one, a key to the species.     

Synthesis and structure of the complex tetra-μ-prolinatodirhodium(II)

The dinuclear Rh^IIRh^II complex with proline [Rh₂(pro₄][NEt₄]₂ was synthesized and its structure studied by means of spectroscopic (IR, EPR and ESCA) and magnetochemical methods. It was shown that two proline molecules serve as bridging ligands, while the other two are only axially coordinated through their N atoms.     

To the knowledge of the fauna of Microlepidoptera (Lepidoptera) of the “Curonian Spit” National Park. II

A list of 243 Microlepidoptera species found in the territory of the Curonian Spit (both in its Russian and Lithuanian parts), in addition to that in the first part of the present publication (Sinev and Shapoval, 2013), is given: Gelechiidae (33), Tortricidae (138), and Crambidae (72 species). 142 species have been collected in the territory of the “Kurshskaya Kosa” National Park, including 49 species new to the Curonian Spit, 30 species new to Kaliningrad Province and 3 species (Scrobipalpa obsoletella, Scrobipalpa pauperella, and Catoptria osthelderi) new to the fauna of Russia.     

Synthesis, Characterization and Antitumor Studies of Mn(II), Ni(II), Cu(II) and Zn(II) Complexes of N-Nicotinoyl-N′-o-Hydroxythiobenzhydrazide

A new ligand N-Nicotinoyl-N-o-hydroxythiobenzhydrazide (H₂Notbh) forms complexes [Mn(Notbh)(H₂O)], [M(Notbh)] [M=Ni(II) Cu(II) and Zn(II)] which were characterized by various physico-chemical techniques. All the metal complexes were observed to inhibit the growth of tumor in vitro, whereas, ligand did not. In vivo administration of these complexes resulted in prolongation of survival of tumor bearing mice. Tumor bearing mice administered with metal complexes showed reversal of tumor growth associated induction of apoptosis in lymphocytes. The paper discusses the possible mechanisms and therapeutic implication of the H₂Notbh and its metal complexes in tumor regression and tumor growth associated immunosuppression.     

On the mechanism of action of thiamin enzymes, Crystal structure of 2-(α-hydroxyethyl)thiamin pyrophosphate (HETPP). Complexes of HETPP with zinc(II) and cadmium(II)

The crystal structure of the 2-(α-hydroxethyl) thiamin pyrophosphate (LH₂) was solved by X-ray diffraction. Crystallographic data: space group F2dd, a=7.922(4) Å, b=33.11(2) Å, c=36.232(10) Å, V=9503(9) ų, z=16. Metal complexes of the general formula K₂{[M(LH)Cl₂]₂} (M=Zn²⁺, Cd²⁺) were isolated from methanolic solutions and characterized by elemental analysis, IR, Raman, and ¹³C CP MAS NMR spectra. They were also characterized by ¹³C NMR, ³¹P NMR, ¹¹³Cd NMR, ES-MS, and ¹H NMR ROESY spectra in D₂O solutions. The data provide evidence for the bonding of the metals to the N(1′) atom of the pyrimidine ring and to the pyrophosphate group. The free ligand and the metal-coordinated ligand adopt the S conformation. Since thiamin cofactor, substrate, and metal ions are present in our system, the extracted results directly refer to thiamin catalysis and possible functional implications are correlated and discussed.     

Synthesis and characterization of β-diketiminate germanium(II) and tin(II) bromides

The equimolar reaction of a β-diketiminate lithium salt LLi(OEt₂) [L = HC(CMeNAr)₂; Ar = 2,6-iPr₂C₆H₃] with either GeBr₂ or SnBr₂ in diethyl ether affords the synthetically useful monomeric β-diketiminate-element halides LGeBr (1) and LSnBr (2), respectively. Both are soluble in hydrocarbon solvents, stable in inert atmosphere, and have been characterized by elemental analysis, NMR spectroscopy, and single-crystal X-ray diffraction analysis.     

Activation of the Si-Si σ-bond of 1,2-divinyldisilane catalyzed by Ru(II) complexes

In the presence of a catalytic amount of RuHCl(CO)(PR₃)_n (R=ⁱPr, n=2; R=Ph, n=3), 1,1,2,2-tetramethyl-1,2-divinyldisilane (1) undergoes unexpected and clean isomerization via the Si-Si bond cleavage to yield a mixture of 6- and 5-membered cyclic compounds, 1,1,4,4-tetramethyl-1,4-disilacyclohex-2-ene and 1,1,2,3,3,-pentamethyl-1,3-disilacyclopent-4-ene, the former being the major product.     

A quantum-chemical study of the binding ability of βXaaHisGlyHis towards copper(II) ion

The present study analyzed binding of Cu²⁺ to tetrapeptides in water solution at several levels of theoretical approximation. The methods used to study the energetic and structural properties of the complexes in question include semiempirical hamiltonians, density functional theory as well as ab initio approaches including electron correlation effects. In order to shed light on the character of interactions between Cu²⁺ and peptides, which are expected to be mainly electrostatic in nature, decomposition of interaction energy into physically meaningful components was applied.     

Detection of low molecular weight copper(II) and zinc(II) binding ligands in ultrafiltered milks—the citrate connection

Low molecular weight zinc(II) and copper(II) binding ligands were detected in ultrafiltered human, bovine, and goat milk by the application of the method of modified gel chromatography. Human milk contains at least three detectable low molecular weight copper binders, whereas bovine and goat milk contain at least two. All three milks show two copper binding peaks with the same elution volumes. Zinc chromatograms were less specific than copper. Zinc showed only a single detectable low molecular weight binding ligand common to all three milks. Elution volumes for both zinc(II) and copper(II) citrate and picolinate systems were measured. Elution volumes of both copper(II) and zinc(II) citrate complexes are identical to elution volumes of an intense peak observed with all three milks; it is reasonable to assume that at least part of this peak corresponds to citrate. Human milk alone has a relatively intense binding peak for copper(II) at the same elution volume as the glutamate complex. Human and goat milk have another low intensity copper(II) binding ligand peak at the same elution volume; a number of amino acid complexes have binding peaks at this position. No peak characteristic of the zinc(II) or copper(II) picolinate systems could be found with any of the milks.     

Mechanism of zinc(II)-promoted amyloid formation: zinc(II) binding facilitates the transition from the partially α-helical conformer to aggregates of amyloid β protein(1–28)

The amyloidoses are a group of disorders characterized by aberrant protein folding and assembly, leading to the deposition of insoluble protein fibrils (amyloid), which provokes cell dysfunction and later cell death. One of the physiologically relevant environmental factors able to affect the conformation and hence the aggregation properties of amyloidogenic proteins/peptides is metal ions. Zn(II) promotes aggregation of most amyloidogenic peptides/proteins in vitro, including amyloid β protein (Aβ), but the underlying mechanism is not known. To better understand this mechanism the present study focused on the partially α-helical conformer, supposed to be an intermediate in Aβ aggregation. This partially α-helical conformer is stabilized by 10–20% 2,2,2-trifluoroethanol (TFE): therefore, the influence of Zn binding on the aggregation of the amylidogenic model peptide Aβ(1–28) (Aβ28) was investigated at different TFE concentrations. The results showed a synergistic effect of Zn(II) and 10% TFE, i.e., that either Zn or 10% TFE accelerated Aβ28 aggregation on its own, but with them together an at least 10 times promotion of Aβ28 aggregation was observed. Further studies by thioflavin T fluorescence spectroscopy, transmission electron microscopy, and circular dichroism (CD) spectroscopy suggested that the aggregates of Zn-Aβ28 formed in 10%TFE contain a β-sheet secondary structure and are more of the amyloid type. CD spectroscopy indicated that Zn binding disrupted partially the α-helical structure of Aβ28 in TFE. Thus, we propose that the promotion of Aβ28 aggregation by Zn is based on the transformation of the partially α-helical conformer (intermediate) towards the β-sheet amyloid structure by a destabilization of the α-helix in the intermediate. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.

Gonadotropin-releasing hormone II (GnRH II) mediates the anorexigenic actions of α-melanocyte-stimulating hormone (α-MSH) and corticotropin-releasing hormone (CRH) in goldfish

Intracerebroventricular (ICV) administration of gonadotropin-releasing hormone II (GnRH II), which plays a crucial role in the regulation of reproduction in vertebrates, markedly reduces food intake in goldfish. However, the neurochemical pathways involved in the anorexigenic action of GnRH II and its interaction with other neuropeptides have not yet been identified. Alpha-melanocyte-stimulating hormone (α-MSH), corticotropin-releasing hormone (CRH) and CRH-related peptides play a major role in feeding control as potent anorexigenic neuropeptides in goldfish. However, our previous study has indicated that the GnRH II-induced anorexigenic action is not blocked by treatment with melanocortin 4 receptor (MC4R) and CRH receptor antagonists. Therefore, in the present study, we further examined whether the anorexigenic effects of α-MSH and CRH in goldfish could be mediated through the GnRH receptor neuronal pathway. ICV injection of the MC4R agonist, melanotan II (80 pmol/g body weight; BW), significantly reduced food intake, and its anorexigenic effect was suppressed by ICV pre-administration of the GnRH type I receptor antagonist, antide (100 pmol/g BW). The CRH-induced (50 pmol/g BW) anorexigenic action was also blocked by treatment with antide. ICV injection of CRH (50 pmol/g BW) induced a significant increase of the GnRH II mRNA level in the hypothalamus, while ICV injection of melanotan II (80 pmol/g BW) had no effect on the level of GnRH II mRNA. These results indicate that, in goldfish, the anorexigenic actions of α-MSH and CRH are mediated through the GnRH type I receptor-signaling pathway, and that the GnRH II system regulates feeding behavior.     

Emission polarization study of the interaction of stellacyanin with tris(2, 2′-bipyridine)osmium(II)

The association of Os(bpy)₃²⁺ and stellacyanin was investigated by measuring the emission polarization of the excited state of the complex [*Os-(bpy)₃²⁺]. At high protein concentration (≥1 mM) Os(bpy)₃²⁺ appears to bind weakly to reduced stellacyanin and to oxidized stellacyanin to a lesser extent. These results are consistent with those obtained in a previous kinetic study on the redox quenching of *Ru(bpy)₃²⁺ by stellacyanin [1]. On excitation at 480 nm, the limiting polarization (P_o) of the *Os(bpy)₃²⁺ emission at 715 nm is 0.100. From P_o we concluded that (1) protein-bound Os(bpy)₃²⁺ has considerable rotational freedom independent of the protein-probe complex, and (2) the emission dipole of *Os(bpy)₃²⁺ is at ∼45° to the C₃ molecular axis.     

Nickel(II)—bleomycin: spectral investigation of the metal binding site

Bleomycin (blm) solutions containing the nickel(II) ion have been investigated through ¹H nmr, ligand field and circular dichroism spectroscopies. It has been found the blm binds the metal ion in a pH dependent fashion. The spectral data are consistent with the presence of at least two species. It is suggested that in the low pH region blm binds to nickel(II) through the β-aminoalanino residue, whereas in the high pH region, the 4-amino-pyrimidine, imidazole, and amido group of β-hydroxy-histidine are also involved in coordination.     

Complexes of Co(II), Ni(II), Cu(II) and Zn(II) with 3′AMP and 2′AMP

Derivatives of Co(II), Ni(II), Cu(II) and Zn(II) with 3′AMP and 2′AMP were synthesized and characterized by IR UV-Vis and fluorescence spectroscopy. There seems to be bonding of the metal ion to the base in all cases. The activation test, using the complexes as allosteric labels, was carried out with rabbit muscle glycogen phosphorylase b, but the enzyme was not activated, confirming that the phosphate group must necessarily be bonded to position 5′ of the ribose in order to activate this enzyme.     

Structural and some medicinal characteristics of the copper(II)–hydroxychloroquine complex

In the first phase of this study, the binding of hydroxychloroquine to the copper(II) cation is examined using liquid chromatography–mass spectrometry (LC–MS), matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS), Fourier transform-ion cyclotron resonance spectrometry (FT-ICR) and nuclear magnetic resonance (¹H and ¹³C NMR) in one and two dimensions. The data suggest the metal–ligand complex is a polarity adaptive molecule. In the second phase of the study, the complexes activity is tested against the National Cancer Institute’s 60 cell line panel. Its anti-cancer activity is compared to quinine, Cu(II)–quinine and hydroxychloroquine. It serves as a base line for future anti-cancer complexes in which hydroxychloroquine is utilized for its ability to impact cell autophagy.     

Dissecting the dimerization motif of Enterococcus hirae’s Zn(II)CopY

The regulation of the copper homeostasis pathway in Enterococcus hirae is conducted through activity of the zinc metalloprotein Zn(II)CopY, which is a Cu(I)-responsive dimeric repressor (Cobine et al., Biochemistry 41:5822-5829, 2002). Its dimerization domain contains a C-terminal cysteine-rich metal-binding motif used for Cu(I) sensing adjacent to an aliphatic-rich repeating sequence, but it is unclear as to which regions contribute most to the interaction. To accomplish this, a synthetically produced CopY construct (CDG) was fused with solubility enhancement tags so the key components of the elements of the aliphatic repeat and metal-binding site could be probed for their dimerization activity. The resultant fusion constructs were tested using two independent methods. Isothermal titration calorimetry, an in vitro technique, was employed to determine dimer affinity thermodynamically. Protein fragment complementation, an in vivo technique, made it possible to rapidly screen homodimeric and heterodimeric complexes within live cells. The combination of in vivo and in vitro studies enabled the identification of CDG sequences that dimerize and sequences that do not, in addition to deciphering relative dimer affinity between all constructs screened. The in vivo technique allowed the formation of heterodimers to be tested for their ability to form specific complexes between dissimilar CDG analogs.