Calcium-induced membrane microdomains trigger plant phospholipase D activity

Plant alpha-type phospholipase D proteins are calcium-dependent, lipolytic enzymes. The morphology of the aggregates of their phospholipid substrate fundamentally defines the interaction between the enzyme and the surface. Here we demonstrate that the Ca(2+)-induced generation of membrane microdomains dramatically activates alpha-type phospholipase D from white cabbage. 500-fold stimulation was observed upon incorporation of 10 mol % 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphate (POPA) into 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) vesicles in the presence of Ca(2+) ions. Enhanced association of PLDalpha2 with phospholipid surfaces containing anionic components was indicated by lag phase analysis and film balance measurements. Differential scanning calorimetry showed that the POPA-specific activation correlates with the phase behavior of the POPC/POPA vesicles in the presence of Ca(2+) ions. We conclude from the results that the Ca(2+)-induced formation of POPA microdomains is the crucial parameter that facilitates the binding of PLD to the phospholipid surface and suggest that this effect serves as a cellular switch for controlling PLD activity.     

The slowly aggregating salmon Calcitonin: a useful tool for the study of the amyloid oligomers structure and activity

Amyloid proteins of different aminoacidic composition share the tendency to misfold and aggregate in a similar way, following common aggregation steps. The process includes the formation of dimers, trimers, and low molecular weight prefibrillar oligomers, characterized by the typical morphology of globules less than 10 nm diameter. The globules spontaneously form linear or annular structures and, eventually, mature fibers. The rate of this process depends on characteristics intrinsic to the different proteins and to environmental conditions (i.e., pH, ionic strength, solvent composition, temperature). In the case of neurodegenerative diseases, it is now generally agreed that the pathogenic aggregates are not the mature fibrils, but the intermediate, soluble oligomers. However, the molecular mechanism by which these oligomers trigger neuronal damage is still unclear. In particular, it is not clear if there is a peculiar structure at the basis of the neurotoxic effect and how this structure interacts with neurons. This review will focus on the results we obtained using salmon Calcitonin, an amyloid protein characterized by a very slow aggregation rate, which allowed us to closely monitor the aggregation process. We used it as a tool to investigate the characteristics of amyloid oligomers formation and their interactions with neuronal cells. Our results indicate that small globules of about 6 nm could be the responsible for the neurotoxic effects. Moreover, our data suggest that the rich content in lipid rafts of neuronal cell plasma membrane may render neurons particularly vulnerable to the amyloid protein toxic effect.     

Is competition between Li+ and Mg2+ the underlying theme in the proposed mechanisms for the pharmacological action of lithium salts in bipolar disorder?

Lithium salts have been in use for the treatment of bipolar disorder for more than 50 years, but their pharmacological mode of action remains a matter of conjecture. Li(+) and Mg(2+) share many physicochemical properties. Not surprisingly, many reported cellular targets for Li(+) action involve Mg(2+)-activated enzymes, which are inhibited by Li(+). In this Account, we describe results from our and other laboratories that suggest that a competition mechanism between Li(+) and Mg(2+) ions for Mg(2+)-binding sites in cellular components is the underlying theme in putative mechanisms of Li(+) action.     

Oxidative Stress and Its Significant Roles in Neurodegenerative Diseases and Cancer

Reactive oxygen and nitrogen species have been implicated in diverse pathophysiological conditions, including inflammation, neurodegenerative diseases and cancer. Accumulating evidence indicates that oxidative damage to biomolecules including lipids, proteins and DNA, contributes to these diseases. Previous studies suggest roles of lipid peroxidation and oxysterols in the development of neurodegenerative diseases and inflammation-related cancer. Our recent studies identifying and characterizing carbonylated proteins reveal oxidative damage to heat shock proteins in neurodegenerative disease models and inflammation-related cancer, suggesting dysfunction in their antioxidative properties. In neurodegenerative diseases, DNA damage may not only play a role in the induction of apoptosis, but also may inhibit cellular division via telomere shortening. Immunohistochemical analyses showed co-localization of oxidative/nitrative DNA lesions and stemness markers in the cells of inflammation-related cancers. Here, we review oxidative stress and its significant roles in neurodegenerative diseases and cancer.     

Molecular Hydrogen Neuroprotection in Post-Ischemic Neurodegeneration in the Form of Alzheimer’s Disease Proteinopathy: Underlying Mechanisms and Potential for Clinical Implementation—Fantasy or Reality?

Currently, there is a lot of public interest in naturally occurring substances with medicinal properties that are minimally toxic, readily available and have an impact on health. Over the past decade, molecular hydrogen has gained the attention of both preclinical and clinical researchers. The death of pyramidal neurons in especially the CA1 area of the hippocampus, increased permeability of the blood-brain barrier, neuroinflammation, amyloid accumulation, tau protein dysfunction, brain atrophy, cognitive deficits and dementia are considered an integral part of the phenomena occurring during brain neurodegeneration after ischemia. This review focuses on assessing the current state of knowledge about the neuroprotective effects of molecular hydrogen following ischemic brain injury. Recent studies in animal models of focal or global cerebral ischemia and cerebral ischemia in humans suggest that hydrogen has pleiotropic neuroprotective properties. One potential mechanism explaining some of the general health benefits of using hydrogen is that it may prevent aging-related changes in cellular proteins such as amyloid and tau protein. We also present evidence that, following ischemia, hydrogen improves cognitive and neurological deficits and prevents or delays the onset of neurodegenerative changes in the brain. The available evidence suggests that molecular hydrogen has neuroprotective properties and may be a new therapeutic agent in the treatment of neurodegenerative diseases such as neurodegeneration following cerebral ischemia with progressive dementia. We also present the experimental and clinical evidence for the efficacy and safety of hydrogen use after cerebral ischemia. The therapeutic benefits of gas therapy open up new promising directions in breaking the translational barrier in the treatment of ischemic stroke.     

Pro-Inflammatory S100A8 and S100A9 Proteins: Self-Assembly into Multifunctional Native and Amyloid Complexes

S100A8 and S100A9 are EF-hand Ca(2+) binding proteins belonging to the S100 family. They are abundant in cytosol of phagocytes and play critical roles in numerous cellular processes such as motility and danger signaling by interacting and modulating the activity of target proteins. S100A8 and S100A9 expression levels increased in many types of cancer, neurodegenerative disorders, inflammatory and autoimmune diseases and they are implicated in the numerous disease pathologies. The Ca(2+) and Zn(2+)-binding properties of S100A8/A9 have a pivotal influence on their conformation and oligomerization state, including self-assembly into homo- and heterodimers, tetramers and larger oligomers. Here we review how the unique chemical and conformational properties of individual proteins and their structural plasticity at the quaternary level account for S100A8/A9 functional diversity. Additional functional diversification occurs via non-covalent assembly into oligomeric and fibrillar amyloid complexes discovered in the aging prostate and reproduced in vitro. This process is also regulated by Ca(2+)and Zn(2+)-binding and effectively competes with the formation of the native complexes. High intrinsic amyloid-forming capacity of S100A8/A9 proteins may lead to their amyloid depositions in numerous ailments characterized by their elevated expression patterns and have additional pathological significance requiring further thorough investigation.     

Upconversion nanoparticles for sensitive and in-depth detection of Cu(2+) ions

Detection of Cu(2+) ions and study of their subcellular distribution in physiological processes are of considerable significance because of their potential environmental and biological applications. Some fluorescence based sensors have been developed for selective detection of Cu(2+) ions, based on organic fluorescent probes that specifically bind to Cu(2+) ions. However, these sensors are not suitable for detection in biological samples due to the short penetration depth of UV/visible light used to excite the fluorescent probes. The use of near-infrared (NIR) light can afford penetration depths of an order of magnitude greater than that of visible light, however, a material that can convert NIR light to visible light is required. A facile method has been developed for in-depth detection of Cu(2+) ions based on fluorescence upconversion. A mesoporous silica shell is coated on upconversion nanoparticles (UCNPs) and a Cu(2+) ion sensitive fluorescent probe, rhodamine B hydrazide, is incorporated into the mesoporous silica. Upon excitation by a NIR light, the UCNPs emit visible light to excite the Cu(2+)-sensitive fluorescent probe. Because of the unique optical properties of UCNPs and their ability to convert NIR light to visible light, this is a feasible method for sensitive and in-depth detection of Cu(2+) ions in a complex biological or environmental sample due to the low autofluorescence and the high penetration depth of NIR light.     

Enhanced sensitivity of a direct SERS technique for Hg(2+) detection based on the investigation of the interaction between silver nanoparticles and mercury ions

Mercury which is a very important pollutant has drawn significant attention in recent research. So far, among the various detection methods, the strategies based on surface-enhanced Raman scattering (SERS) are quite attractive because of the high sensitivity, and especially as it is reported that Hg(2+) can be directly detected by SERS without tagging. However, the procedure for the direct SERS detection of mercury is still unclear with little experimental evidence, limiting further development of Hg(2+) detection by SERS. Herein, we performed a simple method based on SERS for the detection of mercury ions in water without tagging. It is established that in only 2 min, low concentration of Hg(2+) can be recognized based on the decrease of SERS intensity. The detection procedure is investigated by multiple characterizations and the mechanism proven by the obtained data provides a practical way to further improve the sensitivity of the SERS detection. It is demonstrated that the interaction between Hg(2+) and Ag nanoparticles (Ag NPs) could occur in a short time, which includes the complexation of Hg(2+) with citrate and the formation of amalgam due to the reduction of Hg(2+). This interaction influences the surface plasmon resonance (SPR) property of Ag NPs and thereby decays the electromagnetic enhancement of Ag NPs; meanwhile the interaction also causes the zeta potential decrease of Ag NPs and accordingly affects the adsorption of Raman reporter molecules on the surface of Ag NPs. Therefore, the weakness of SERS intensity in the presence of Hg(2+) should be mainly attributed to the interaction between Hg(2+) and Ag NPs. From the mechanism demonstrated, it can be speculated that using fewer Ag NPs in the detection could improve the sensitivity, because at low Hg(2+) concentration the interaction becomes stronger since every Ag nanoparticle acts with more Hg(2+) ions. Accordingly, we establish that 90.9 pM (18.2 ppt) Hg(2+) is detected in 18 μM Ag NPs, which is much lower than that in reported papers.     

Copper binding in the prion protein

A conformational change of the prion protein is responsible for a class of neurodegenerative diseases called the transmissible spongiform encephalopathies that include mad cow disease and the human afflictions kuru and Creutzfeldt-Jakob disease. Despite the attention given to these diseases, the normal function of the prion protein in healthy tissue is unknown. Research over the past few years, however, demonstrates that the prion protein is a copper binding protein with high selectivity for Cu(2+). The structural features of the Cu(2+) binding sites have now been characterized and are providing important clues about the normal function of the prion protein and perhaps how metals or loss of protein function play a role in disease. The link between prion protein and copper may provide insight into the general, and recently appreciated, role of metals in neurodegenerative disease.     

Trends in the Molecular Pathogenesis and Clinical Therapeutics of Common Neurodegenerative Disorders

The term neurodegenerative disorders, encompasses a variety of underlying conditions, sporadic and/or familial and are characterized by the persistent loss of neuronal subtypes. These disorders can disrupt molecular pathways, synapses, neuronal subpopulations and local circuits in specific brain regions, as well as higher-order neural networks. Abnormal network activities may result in a vicious cycle, further impairing the integrity and functions of neurons and synapses, for example, through aberrant excitation or inhibition. The most common neurodegenerative disorders are Alzheimer’s disease, Parkinson’s disease, Amyotrophic Lateral Sclerosis and Huntington’s disease. The molecular features of these disorders have been extensively researched and various unique neurotherapeutic interventions have been developed. However, there is an enormous coercion to integrate the existing knowledge in order to intensify the reliability with which neurodegenerative disorders can be diagnosed and treated. The objective of this review article is therefore to assimilate these disorders’ in terms of their neuropathology, neurogenetics, etiology, trends in pharmacological treatment, clinical management, and the use of innovative neurotherapeutic interventions.     

Dissimilar roles of the four conserved acidic residues in the thermal stability of poly(a)-specific ribonuclease

Divalent metal ions are essential for the efficient catalysis and structural stability of many nucleotidyl-transfer enzymes. Poly(A)-specific ribonuclease (PARN) belongs to the DEDD superfamily of 3'-exonucleases, and the active site of PARN contains four conserved acidic amino acid residues that coordinate two Mg(2+) ions. In this research, we studied the roles of these four acidic residues in PARN thermal stability by mutational analysis. It was found that Mg(2+) significantly decreased the rate but increased the aggregate size of the 54 kDa wild-type PARN in a concentration-dependent manner. All of the four mutants decreased PARN thermal aggregation, while the aggregation kinetics of the mutants exhibited dissimilar Mg(2+)-dependent behavior. A comparison of the kinetic parameters indicated that Asp28 was the most crucial one to the binding of the two Mg(2+) ions, while metal B might be more important in PARN structural stability. The spectroscopic and aggregation results also suggested that the alterations in the active site structure by metal binding or mutations might lead to a global conformational change of the PARN molecule.     

Copper and Zinc Binding to Amyloid‐β: Coordination,Dynamics, Aggregation,Reactivity and Metal‐Ion Transfer

The metal ions copper, zinc and iron have been shown to be involved in Alzheimer's disease (AD). Cu, Zn and Fe ions are proposed to be implicated in two key steps of AD pathology: 1) aggregation of the peptide amyloid‐β (Aβ), and 2) production of reactive oxygen species (ROS) induced by Aβ. There is compelling evidence that Cu and Zn bind directly to Aβ in AD. This formation of Cu/Zn–Aβ complexes is thought to be aberrant as they have been detected only in AD, but not under healthy conditions. In this context, the understanding of how these metal ions interact with Aβ, their influence on structure and oligomerization become an important issue for AD. Moreover, the mechanism of ROS production by Cu–Aβ in relation to its aggregations state, as well as the metal‐transfer reaction from and to Aβ are crucial in order to understand why Aβ oligomers are highly toxic and why Aβ seems to bind Cu and Zn only in AD.     

Cul-Cu_2O-WO_3(MoO_3)快离子导电玻璃的ESR研究

本文报道了CuI-Cu_2O-Wo_3(MoO_3)快离子Cu~+导电玻璃在室温下的顺磁共振吸收谱。实验结果表明玻璃的顺磁共振吸收主要由W~(5+),W(3+),和Mo~(5+),Mo~(3+)以及Cu~(2+)离子引起。W~(5+),W~(3+),Mo~(5+)和Mo~(3+)离子均处于对称的四面体中,且W~(3+)和Mo~(3+)离子表现出强烈的d~6离子的高自旋态的吸收特征。Cu~(2+)离子的吸收谱则可用四方畸变的八面体场作用来解释。铜含量较低时吸收以W~(5+),W~(8+),Mo~(5+),Mo~(8+)为主,随着铜含量的增加,Cu~(2+)离子吸收变得显著。     

Role of the Ubiquitin System in Stress Granule Metabolism

Eukaryotic cells react to various stress conditions with the rapid formation of membrane-less organelles called stress granules (SGs). SGs form by multivalent interactions between RNAs and RNA-binding proteins and are believed to protect stalled translation initiation complexes from stress-induced degradation. SGs contain hundreds of different mRNAs and proteins, and their assembly and disassembly are tightly controlled by post-translational modifications. The ubiquitin system, which mediates the covalent modification of target proteins with the small protein ubiquitin (‘ubiquitylation’), has been implicated in different aspects of SG metabolism, but specific functions in SG turnover have only recently emerged. Here, we summarize the evidence for the presence of ubiquitylated proteins at SGs, review the functions of different components of the ubiquitin system in SG formation and clearance, and discuss the link between perturbed SG clearance and the pathogenesis of neurodegenerative disorders. We conclude that the ubiquitin system plays an important, medically relevant role in SG biology.     

通过络合方法强化电渗析对二价同电荷重金属离子分离效果的初步研究

对二价过渡金属离子选择适当的配位体以形成不同的络合物,通过改变电荷,离子在膜内的淌度此及分配系数,或络合物的稳定常数的差异来达到同电荷离子彼此分离的目的。研究结果表明,对形成铜氨络离子—镁离子溶液体系和铜乙二胺络离子—镁离子溶液体系,采用常规电渗析方法进行分离,其T_(Mg)~(Cu)值较常规溶液体系分别提高近一倍和25%;对铜—镁混合溶液和钴—镍混合溶液采用EDTA 络合剂和特殊电渗析工艺,可使T_(Mg)~(Cu)→0或T_(Nl)~(Co)→∞,达到完全分离。本研究开发的分离同电荷离子的电渗析新工艺,对于开拓电渗析技术的新应用领域具有一定的指导意义。     

Solution Equilibria between Aluminum(III) Ion and L-histidine or L-tyrosine

Toxic effects due to high aluminum body loads were observed in a number of conditions following ingestion of Al-containing antacids. Bio-availability of aluminum depends not only on the solubility of the ingested salt but also on the physico-chemical properties of the soluble Al complexes formed in body fluids. Amino acids may, upon interaction with Al-salts, form absorbable Al-complexes. Hence, complex formation equilibria between Al(3+) and either, L- histidine or L-tyrosine were studied by glass electrode potentiometric (0.1 mol/L LiCl ionic medium, 298 K), proton NMR and uv spectrophotometric measurements. Non linear least squares treatment of the potentiometric data indicates that in the concentration ranges: 0.5相似文献   

Stimulation of Phospholipase A2 by Toxic Main Group Heavy Metals: Partly Dependent on G-proteins?

Organometals induce platelet aggregation and inorganic metal ions such as Cd(2+) or Pb(2+) sensitise human blood platelets to aggregating agents and this action is associated with the liberation of arachidonic acid and eicosanoid formation. The same mechanism is observed using human leukaemia cells (HL-60) when treated with MeHgCl or Et(3)PbCl. The fatty acid liberation within human platelets and HL-60 cells could only be inhibited with phospholipase A(2) inhibitors of different specificity.Preincubation of the cells with pertussis toxin reduces the activation induced by Et(3)PbCl to a great extent. The non-catalytic B subunit, that only mediates the binding of the toxin to the cell membranes, has no effect at all. When summarised, these results suggest that one possible mechanism for the stimulation of phospholipase A(2) by Et(3)PbCl functions via a G-protein dependent pathway.