RESUMO
α-Synuclein (αSyn) constitutes the main protein component of Lewy bodies, which are the pathologic hallmark in Parkinson's disease. αSyn is unstructured in solution but the interaction of αSyn with lipid membrane modulates its conformation by inducing an α-helical structure of the N-terminal region. In addition, the interaction with metal ions can trigger αSyn conformation upon binding and/or through the metal-promoted generation of reactive oxygen species which lead to a cascade of structural alterations. For these reasons, the ternary interaction between αSyn, copper, and membranes needs to be elucidated in detail. Here, we investigated the structural properties of copper-αSyn binding through NMR, EPR, and XAS analyses, with particular emphasis on copper(I) coordination since the reduced state is particularly relevant for oxygen activation chemistry. The analysis was performed in different membrane model systems, such as micellar sodium dodecyl sulfate (SDS) and unilamellar vesicles, comparing the binding of full-length αSyn and N-terminal peptide fragments. The presence of membrane-like environments induced the formation of a copper:αSyn = 1:2 complex where Cu+ was bound to the Met1 and Met5 residues of two helical peptide chains. In this coordination, Cu+ is stabilized and is unreactive in the presence of O2 in catechol substrate oxidation.
Assuntos
Doença de Parkinson , alfa-Sinucleína , Humanos , alfa-Sinucleína/metabolismo , Cobre/química , Doença de Parkinson/metabolismo , Peptídeos/metabolismo , OxirreduçãoRESUMO
The effect of Cu2+ on α-synuclein (AS) aggregation is important because clinical studies of patients with Parkinson's disease have shown elevated levels of Cu2+ in the cerebrospinal fluid. So far, the molecular architectures of Cu2+-AS fibril complexes at atomic resolution are unknown. The current work identifies for the first time that His50 cannot bind Cu2+ ions in mature fibrils. Moreover, it shows hopping of Cu2+ ions between residues in AS fibrils and changes in the Cu2+ coordination mode in Cu2+ ions that bind in the termini of AS. The current study combines extensive experimental techniques, density functional theory calculations, and computational modeling tools to provide a complete description of the Cu2+ binding site in AS fibrils. Our findings illustrate for the first time the specific interactions between Cu2+ ions and AS fibrils, suggesting a new mechanistic perspective on the effect of Cu2+ ions on AS aggregation.
RESUMO
Silver is a non-essential element with promising antimicrobial and anticancer properties. This work is a detailed summary of the newest findings on the bioinorganic chemistry of silver, with a special focus on the applications of Ag+ complexes and nanoparticles. The coordination chemistry of silver is given a reasonable amount of attention, summarizing the most common silver binding sites and giving examples of such binding motifs in biologically important proteins. Possible applications of this metal and its complexes in medicine, particularly as antibacterial and antifungal agents and in cancer therapy, are discussed in detail. The most recent data on silver nanoparticles are also summarized.
Assuntos
Anti-Infecciosos/química , Antineoplásicos/química , Complexos de Coordenação/química , Prata/química , Sequência de Aminoácidos , Aminoácidos/química , Anti-Infecciosos/efeitos adversos , Anti-Infecciosos/farmacologia , Antineoplásicos/efeitos adversos , Antineoplásicos/farmacologia , Complexos de Coordenação/efeitos adversos , Complexos de Coordenação/farmacologia , Descoberta de Drogas/métodos , Humanos , Íons/química , Nanopartículas Metálicas/química , Estrutura Molecular , Ligação Proteica , Relação Estrutura-AtividadeRESUMO
BACKGROUND: The understanding of the bioinorganic and coordination chemistry of metalloproteins containing unusual poly-Xaa sequences, in which a single amino acid is repeated consecutively, is crucial for describing their metal binding-structure-function relationship, and therefore also crucial for understanding their medicinal potential. To the best of our knowledge, this is the first systematic review on metal complexes with polyXaa sequences. METHODS: We performed a thorough search of high quality peer reviewed literature on poly-Xaa type of sequences in proteins, focusing on their biological importance and on their interactions with metal ions. RESULTS: 228 papers were included in the review. More than 70% of them discussed the role of metal complexes with the studied types of sequences. In this work, we showed numerous medically important and chemically fascinating examples of possible 'poly-Xaa' metal binding sequences. CONCLUSION: Poly-Xaa sequences, in which a single amino acid is repeated consecutively, are often not only tempting binding sites for metal ions, but very often, together with the bound metal, serve as structure determinants for entire proteins. This, in turn, can have consequences for the whole organism. Such sequences in bacterial metal chaperones can be a possible target for novel, antimicrobial therapeutics.
Assuntos
Aminoácidos/química , Complexos de Coordenação/química , Proteínas/química , Sequência de Aminoácidos , Humanos , Íons/químicaRESUMO
Hemopressin is a neuropeptide, derived from the degradation of the α(1)-chain of hemoglobin, and possesses several pharmacologic properties, such as the ability to block cannabinoid CB1 receptor activity, to cause dose-dependent hypotension and to inhibit food intake. Actually, human hemopressin (PVNFKLLSH) is only the precursor of a class of longer peptides, called "Pepcans", which bear additional residues at their amino-terminus and possess slightly different chemical and biological properties with respect to hemopressin. The presence of a histidyl residue and the free terminal amine imparts to hemopressin and its derivatives good binding properties towards transition metal ions. In this paper, we present a wide investigation on the complex-formation equilibria of human hemopressin and three analogues towards the Cu(ii) and Ni(ii) ions. The study showed that the main coordination site is always the amino terminus (if not protected), while the C-terminal histidine acts only as an anchoring site for the metal ions at acidic pH, with the formation of a macrochelate complex. The presence of additional residues in N-terminal position produces significant differences in the protonation and complex-formation behaviors of these peptides, which can be explained in terms of charge of the ligand and coordination environment. Although the participation of metal ions in the biological activity of hemopressin and Pepcans has not yet been demonstrated, the data reported here can help to shed light on the mechanisms governing the action of these neuropeptides in vivo.
Assuntos
Cobre/química , Hemoglobinas/química , Níquel/química , Fragmentos de Peptídeos/química , Histidina/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Íons/química , Ligantes , Espectroscopia de Ressonância Magnética , Ligação Proteica , Prótons , Receptor CB1 de Canabinoide/antagonistas & inibidores , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
The Eighth Central European Conference "Chemistry towards Biology" was held in Brno, Czech Republic, on August 28-September 1, 2016 to bring together experts in biology, chemistry and design of bioactive compounds; promote the exchange of scientific results, methods and ideas; and encourage cooperation between researchers from all over the world. The topics of the conference covered "Chemistry towards Biology", meaning that the event welcomed chemists working on biology-related problems, biologists using chemical methods, and students and other researchers of the respective areas that fall within the common scope of chemistry and biology. The authors of this manuscript are plenary speakers and other participants of the symposium and members of their research teams. The following summary highlights the major points/topics of the meeting.
Assuntos
Química Farmacêutica/métodos , Proteínas/química , Sistemas de Liberação de Medicamentos , Desenho de Fármacos , Epigênese Genética , Relação Estrutura-Atividade , Biologia de SistemasRESUMO
The basic knowledge about biological inorganic chemistry, thermodynamics and metal binding sites of metalloproteins is crucial for the understanding of their metal binding-structure-function relationship. Metal-peptide complexes are useful and commonly used models of metal-enzyme active sites, among which copper and zinc models are one of the most extensively studied. HENRYK is a peptide sequence present in numerous proteins, and serves as a potentially tempting binding site for Cu2+ and Zn2+. Maybe more importantly, HENRYK also happens to be the first name of our group leader. The results of this work, which, at the first glance, might seem to be a 'chemical scrabble', went far beyond our expectations and surprised us with a novel, uncommon behavior of a Cu2+ complex with a peptide with a histidine in position one. At low pH, the binding is a typical histamine-like coordination, but with the increase of pH, the imidazole nitrogen is moved to the axial position and replaced with an amide; at basic pH, the binding mode is a {NH2, 3N-} one in the equatorial plane. It is important to note, that no dimeric species are formed in between. Such binding is thermodynamically much more stable than a simple complex with histamine, and quite comparable to complexes with several possible imidazole anchoring sites.
Assuntos
Complexos de Coordenação/química , Cobre/química , Peptídeos/química , Zinco/química , Histamina/química , Concentração de Íons de HidrogênioRESUMO
The zinc binding loop domain of the HypA protein of Helicobacter pylori consists of two CXXC motifs with flanking His residues. These motifs bind metal ions, and thus they are crucial for the functioning of the whole protein. The N-terminal site, where His is separated from CXXC by Ser residue is more effective in binding Zn(2+) and Ni(2+) ions than the C-terminal site, in which His is adjacent to the CXXC motif. Studies on various modifications of the peptide sequence within the Ac-ELECKDCSHVFKPNALDYGVCEKCHS-NH2 loop show the role of the residues in the linker between the CXXC motifs and the effect of the length of the linker on the stability of the complexes it forms with Zn(2+), Cd(2+) and Ni(2+) ions. The proline residue in the linker between the two CXXC binding sites plays a distinct role in the metal ion binding ability of the loop, lowering the efficacy of the metal ion coordination. The deletion of the aliphatic residues from the linker between the CXXC motifs remarkably improves the binding efficacy of the loop.
Assuntos
Proteínas de Bactérias/química , Cádmio/química , Proteínas de Transporte/química , Níquel/química , Peptídeos/química , Zinco/química , Sítios de Ligação , Histidina/química , Metalochaperonas , Ligação Proteica , Estrutura Terciária de ProteínaRESUMO
The acid-base properties and the Cu(II) binding processes of methotrexate (MTX) were characterized by selected spectroscopic techniques and potentiometric measurements. The pH titration data showed that MTX behaves as a triprotic ligand. The deprotonation constants were determined for α-COOH and γ-COOH groups and (N1)H+ from the pteridine ring. Taking all the obtained results into consideration, a coordination pattern was proposed. The DNA-cleaving activity and reactive oxygen species (ROS) generation were investigated for both MTX and the Cu(II)-MTX system. The complex displayed a promising nuclease activity toward plasmid DNA in the presence of hydrogen peroxide. Interestingly, the induction of ROS, such as hydroxyl radicals, superoxide anions or singlet oxygen, was excluded and a different mechanism of DNA degradation was proposed. As MTX is now commonly used in anticancer therapy i.e. against lung cancer, basic cell-based studies were carried out to establish if its Cu(II) complex exhibits higher cytotoxic properties than the ligand alone. Activities of both compounds were also tested against colon carcinoma. Moreover, the determined values of IC50 were confronted with the cytotoxic activity of cisplatin.