The effect of a membrane-mimicking environment on the interactions of Cu2+ with an amyloidogenic fragment of chicken prion protein

Prion proteins (PrP) from different species have the ability to tightly bind Cu2+ ions

The Eighth Central European Conference "Chemistry towards Biology": snapshot

The Eighth Central European Conference "Chemistry towards Biology" was held in Brno, Czech Republic, on 28 August – 1 September 2016The Eighth Central European Conference "Chemistry towards Biology" was held in Brno, Czech Republic, on 28 August-1 September 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

Zinc(II)—The Overlooked Éminence Grise of Chloroquine’s Fight against COVID-19?

The authors would like to thank Agnieszka Michalczuk for providing us with her artistic vision of SARS-CoV-2.Zn(II) is an inhibitor of SARS-CoV-2′s RNA-dependent RNA polymerase, and chloroquine and hydroxychloroquine are Zn(II) ionophores–this statement gives a curious mind a lot to think about. We show results of the first clinical trials on chloroquine (CQ) and hydroxychloroquine (HCQ) in the treatment of COVID-19, as well as earlier reports on the anticoronaviral properties of these two compounds and of Zn(II) itself. Other FDA-approved Zn(II) ionophores are given a decent amount of attention and are thought of as possible COVID-19 therapeutics.National Science Center, Poland UMO-2017/26/A/ST5/00363 UMO-2017/26/A/ST5/0036

Influence of membrane environments and copper ions on the structural features of amyloidogenic proteins correlated to neurodegeneration

Amyloidogenic proteins are associated with severe neurodegenerative disorders afflicting millions of people worldwide. The main hallmarks of these diseases are the presence of amyloid plaques in the brain, primarily formed by fibrils of misfolded cellular proteins. The process leading to protein aggregation, is far from completely understood. However, it is well accepted that many factors are able to influence the morphology and kinetics of amyloids formation. In the last years, a plethora of studies have suggested that the aggregation process is greatly influenced by the interactions of these proteins with copper and membranes. Amyloidogenic proteins undergo large conformational changes in the presence of membranes and show the presence of stable α helix structures in the presence of high concentrations of unilamellar vesicles or detergent micelles. The regions involved in the helicoidal rearrangements are those which are critical for protein aggregation. In addition, the α helix structuring of amyloidogenic proteins may strongly affect copper binding, in terms of donor atoms and complex stability since metal binding domains are often located near regions experiencing transitions from random-coil to α helix conformations

Investigation of metal interactions with YrpE protein of Bacillus subtilis by a polyhistidine peptide model

The use of model peptides that can simulate the behaviour of a protein domain is a very successful analytical method to study the metal coordination sites in biological systems. Here we study zinc and copper binding ability of the sequence HTHEHSHDHSHAH, which serves as model for the metal interactions with YrpE, a putative metal-binding protein of the ZinT family identified in Bacillus subtilis. Compared to other ZinT proteins secreted by Gram-negative bacteria, the metal-coordination properties of YrpE N-terminal histidine-rich domain have not been yet characterized. Different independent analytical methods, aimed at providing information on the stability and structure of the formed species, have been employed, including potentiometric titrations, electrospray ionization mass spectrometry, UV-Vis spectrophotometry, circular dichroism and electron paramagnetic resonance spectroscopy. The obtained speciation models and equilibrium constants allowed to compare the metal-binding ability of the investigated polyhistidine sequence with that of other well-known histidine-rich peptides. Our thermodynamic results revealed that the YrpE domain HTHEHSHDHSHAH forms more stable metal complexes than other His-rich domains of similar ZinT proteins. Moreover, the studied peptide, containing the alternated (-XH-)n motif, proved to be even more effective than the His6-tag (widely used in immobilized metal ion affinity chromatography) in binding zinc ions

Novel Perspective on Alzheimer's Disease Treatment: Rosmarinic Acid Molecular Interplay with Copper(II) and Amyloid beta

Alzheimer’s disease is a severe disorder that affects millions of people worldwide. It is a very debilitating disease with no cure at the moment. The necessity of finding an effective treatment is very demanding, and the entire scientific community is putting in a lot of effort to address this issue. The major hallmark of Alzheimer’s disease is the presence of toxic aggregated species in the brain, impaired metal homeostasis, and high levels of oxidative stress. Rosmarinic acid is a well-known potent antioxidant molecule, the efficacy of which has been proved both in vitro and in vivo. In this study, we investigated the possible role played by rosmarinic acid as a mediator of the copper(II)-induced neurotoxicity. Several spectroscopic techniques and biological assays were applied to characterize the metal complexes and to evaluate the cytotoxicity and the mutagenicity of rosmarinic acid and its Cu(II) complex. Our data indicate that rosmarinic acid is able to interfere with the interaction between amyloid β and Cu(II) by forming an original ternary association

Impact of SDS surfactant on the interactions of Cu(2+) ions with the amyloidogenic region of human prion protein

Prion diseases, known as Transmissible Spongiform Encephalopathies (TSEs), are a group of fatal neuronal, and to some extent infectious disorders, associated with a pathogenic protein agent called prion protein (PrP). The human prion protein (hPrP) fragment encompassing the 91-127 region, also known as the amyloidogenic domain, comprises two copper-binding sites corresponding to His-96 and His-111 residues that act as anchors for Cu(2+) binding. In this work, we investigated Cu(2+) interaction with hPrP91-127 in the presence of the anionic surfactant sodium dodecyl sulfate (SDS), which induces a partial α-helix folding of the peptide. Our data indicate that the Cu(2+) coordination ability of the amyloidogenic fragment in the presence of SDS micelles is significantly different to that observed in aqueous solution. This is mainly due to the fact that SDS micelles strongly stabilize the formation of the α-helical structure of the peptide backbone, which is well conserved also upon Cu(2+) binding, contrary to the random coil conformation mainly assumed by hPrP91-127 in aqueous solutions. Potentiometric and spectroscopic studies clearly indicate that in the case of SDS containing solutions, Cu(2+) ions coordinate simultaneously to both imidazoles, while in the case of water solutions, metal ion coordination involves only a single His side chain, which individually acts as an independent Cu(2+) anchoring site

Specific binding modes of Cu(I) and Ag(I) with neurotoxic domain of the human prion protein

Prion diseases are neurodegenerative disorders associated with a conformational change of the normal cellular isoform of the prion protein (PrPC) to an abnormal scrapie isoform (PrPSc). human prion protein (hPrPC) is able to bind up to six Cu(II) ions. Four of them are distributed in the octarepeat domain, containing four tandem-repetitions of the sequence PHGGGWGQ. Immediately outside the octarepeat domain, in so called PrP amyloidogenic region, two additional and independent Cu(II) binding sites, encompassing His96 and His111 residues, respectively, are present. Considering the potential involvement of PrP in cellular redox homeostasis, investigations on Cu(I)-PrP interaction might be also biologically relevant. Interestingly, the amyloidogenic fragment of PrP contains a -M(X)nM- motif, known to act as Cu(I) binding site in different proteins. In order to shed more light on this issue, copper(I) and silver(I) interactions with model peptides derived from that region were analyzed. The results of our studies reveal that both metal ions are anchored to two thioether sulfurs of Met109 and Met112, respectively. Subsequent metal interaction and coordination to His96 and His111 imidazoles are primarily found for Cu(I) at physiological pH. Metal binding was also investigated in the presence of negatively charged micelles formed by the anionic surfactant, sodium dodecyl sulfate (SDS). Our results strongly support that metal binding mode strongly depends on the protein backbone structure. In particular we show that α-helix structuring of the amyloid PrP domain influences both the metal coordination sphere and the binding affinity

The interaction of antimicrobal peptides with metal ions : the relationship between coordination chemistry, structure, thermodynamics and mode of action

Increasing bacterial and fungal drug resistance makes novel, effective antimicrobial treatments actively sought. Because of the general lack of resistance towards antimicrobial peptides (AMPs), they are being relied on as a novel class of therapeutics aimed to conquer drug-resistant bacteria and fungi. There are numerous ways in which AMPs might interact with pathogens, such as membrane disruption, production of reactive oxygen species, inhibition of cell wall, nucleic acid and protein synthesis or by the withdrawal of essential metal ions. Biologically indispensable metal ions, such as Zn(II) and Cu(II), which are the key players of this project, have a dual effect on the activity of antimicrobial peptides: (i) AMPs bind them, so that microbes cannot get enough metals essential for their life and virulence (withdrawal of metal ions, nutritional immunity) or (ii) AMPs need the given metal ion as a booster of their antimicrobial activity (metal ions affect the AMP charge and/or structure). In this chapter, we discuss the impact of the coordination of Cu(II) and Zn(II) to several antimicrobial peptides, focusing on the thermodynamics, structure and coordination chemistry. The comparison of these data to the outcome of biological growth studies (determination of minimal inhibitory concentration (MIC) of metalAMP complexes and their derivatives allows to draw conclusions about the relationship between the metal-antimicrobial peptide complex structure, stability mode of action and efficacy. In the nearest future, the most efficient complexes may serve as templates for a rational design of novel, more potent AMP-based therapeutics. Further improvement can be reached through the modification of the most promising AMP complexes using (i) specifically targeted antimicrobial peptides, in which the AMP will be covalently linked to a targeting peptide (Figure 1) or (ii) chimeric compounds comprising AMPs bound to conventional antimicrobials or peptidomimetic modifications (Figure 2)