Molecular Dynamics Simulation of Nucleation and Growth of a Binary Quasicrystal

Abstract

The quasicrystal structure is considered to be a new type of ordered phase because its Fourier transform has Laue spots with icosahedral symmetry, which is inconsistent with crystal structure. Computer simulation of the formation process of a quasicrystal was performed by the molecular dynamics method. On the basis of the Strandburg type of quasicrystal model, we developed an algorithm of the formation process of binary quasicrystal reflecting the procedure as realistically as possible. The Fourier transform of some of the obtained structures has shown decagonal symmetry although the spots are rather diffused. It has been shown that the potential parameter and experimental condition should be limited to produce a perfect quasicrystal structure.     

A model of cell cycle behavior dominated by kinetics of a pathway stimulated by growth factors

A modified version of a previously developed mathematical model [Obeyesekere et al., Cell Prolif. (1997)] of the G1-phase of the cell cycle is presented. This model describes the regulation of the G1-phase that includes the interactions of the nuclear proteins, RB, cyclin E, cyclin D, cdk2, cdk4 and E2F. The effects of the growth factors on cyclin D synthesis under saturated or unsaturated growth factor conditions are investigated based on this model. The solutions to this model (a system of nonlinear ordinary differential equations) are discussed with respect to existing experiments. Predictions based on mathematical analysis of this model are presented. In particular, results are presented on the existence of two stablesolutions, i. e., bistability within the G1-phase. It is shown that this bistability exists under unsaturated growth factor concentration levels. This phenomenon is very noticeable if the efficiency of the signal transduction, initiated by the growth factors leading to cyclin D synthesis, is low. The biological significance of this result as well as possible experimental designs to test these predictions are presented.     

A Clinical Prediction Model to Distinguish Maturity-Onset Diabetes of the Young From Type 1 and Type 2 Diabetes in the Chinese Population

ObjectiveGenetic detection for the diagnosis of maturity-onset diabetes of the young (MODY) in China has low sensitivity and specificity. Better gene detection is urgently needed to distinguish testing subjects. We proposed to use numerous and weighted clinical traits as key indicators for reasonable genetic testing to predict the probability of MODY in the Chinese population.MethodsWe created a prediction model based on data from 306 patients, including 140 patients with MODY, 84 patients with type 1 diabetes (T1D), and 82 patients with type 2 diabetes (T2D). This model was evaluated using receiver operating characteristic curves.ResultsCompared with patients with T1D, patients with MODY had higher C-peptide levels and negative antibodies, and most patients with MODY had a family history of diabetes. Different from T2D, MODY was characterized by lower body mass index and younger diagnostic age. A clinical prediction model was established to define the comprehensive probability of MODY by a weighted consolidation of the most distinguishing features, and the model showed excellent discrimination (areas under the curve of 0.916 in MODY vs T1D and 0.942 in MODY vs T2D). Further, high-sensitivity C-reactive protein, glycated hemoglobin A1c, 2-h postprandial glucose, and triglyceride were used as indicators for glucokinase-MODY, while triglyceride, high-sensitivity C-reactive protein, and hepatocellular adenoma were used as indicators for hepatocyte nuclear factor 1-α MODY.ConclusionWe developed a practical prediction model that could predict the probability of MODY and provide information to identify glucokinase-MODY and hepatocyte nuclear factor 1-α MODY. These results provide an advanced and more reasonable process to identify the most appropriate patients for genetic testing.     

A toolset to study functions of Cytosolic non-specific dipeptidase 2 (CNDP2) using Drosophila as a model organism

Background

Expression of the CNDP2 gene is frequently up- or down-regulated in different types of human cancers. However, how the product of this gene is involved in cell growth and proliferation is poorly understood. Moreover, our knowledge of the functions of the CNDP2 orthologs in well-established model organisms is scarce. In particular, the function of the D. melanogaster ortholog of CNDP2, encoded by the CG17337 gene (hereafter referred to as dCNDP2), is still unknown.

Results

This study was aimed at developing a set of genetic and molecular tools to study the roles of dCNDP2. We generated a dCNDP2 null mutation (hereafter ∆dCNDP2) using CRISPR/Cas9-mediated homologous recombination (HR) and found that the ∆dCNDP2 mutants are homozygous viable, morphologically normal and fertile. We also generated transgenic fly lines expressing eGFP-tagged and non-tagged dCNDP2 protein, all under the control of the UAS promoter, as well as polyclonal antibodies specific to dCNDP2. Using these tools, we demonstrate that only one of the two predicted dCNDP2 isoforms is expressed throughout the different tissues tested. dCNDP2 was detected in both the cytoplasm and the nucleus, and was found to be associated with multiple sites in the salivary gland polytene chromosomes.

Conclusions

The dCNDP2 gene is not essential for fly viability under standard laboratory conditions. The subcellular localization pattern of dCNDP2 suggests that this protein might have roles in both the cytoplasm and the nucleus. The genetic and molecular tools developed in this study will allow further functional characterization of the conserved CNDP2 protein using D. melanogaster as a model system.

     

Theoretical Population Dynamics Model of a Genetically Transmitted Disease: Sickle-Cell Anaemia

We formulate a realistic demographic model that captures the pattern of inheritance of the S gene, which is responsible for the most common genetic defect, namely, sickle-cell anaemia (SCA), using general pair formations. The model equation is implicitly solved via the Laplace transform technique, while the existence of a unique solution is proved by applying the contraction mapping principle. One of the main results is the boundedness of the solution. A fundamental reason for the persistence of SCA is probably due to the role played by the selective advantage of the abnormal S gene over the normal haemoglobin A in tropical regions, and the fact that carriers are more fertile and survive longer (a property known as hybrid vigor), because they are essentially asymptomatic. We also discuss possible public health policies. 2000 MS Classification: 92D25 · 92D40 · 34C60 · 35F10     

Proliferation and tenogenic differentiation of bone marrow mesenchymal stem cells in a porous collagen sponge scaffold

BACKGROUND Collagen is one of the most commonly used natural biomaterials for tendon tissue engineering.One of the possible practical ways to further enhance tendon repair is to combine a porous collagen sponge scaffold with a suitable growth factor or cytokine that has an inherent ability to promote the recruitment,proliferation,and tenogenic differentiation of cells.However,there is an incomplete understanding of which growth factors are sufficient and optimal for the tenogenic differentiation of rat bone marrow mesenchymal stem cells(BMSCs)in a collagen sponge-based 3D culture system.AIM To identify one or more ideal growth factors that benefit the proliferation and tenogenic differentiation of rat BMSCs in a porous collagen sponge scaffold.METHODS We constructed a 3D culture system based on a type I collagen sponge scaffold.The surface topography of the collagen sponge scaffold was observed by scanning electron microscopy.Primary BMSCs were isolated from Sprague-Dawley rats.Cell survival on the surfaces of the scaffolds with different growth factors was assessed by live/dead assay and CCK-8 assay.The mRNA and protein expression levels were confirmed by quantitative real-time polymerase chain reaction and Western blot,respectively.The deposited collagen was assessed by Sirius Red staining.RESULTS Transforming growth factorβ1(TGF-β1)showed great promise in the tenogenic differentiation of BMSCs compared to growth differentiation factor 7(GDF-7)and insulin-like growth factor 1(IGF-1)in both the 2D and 3D cultures,and the 3D culture enhanced the differentiation of BMSCs into tenocytes well beyond the level of induction in the 2D culture after TGF-β1 treatment.In the 2D culture,the proliferation of the BMSCs showed no significant changes compared to the control group after TGF-β1,IGF-1,or GDF-7 treatment.However,TGF-β1 and GDF-7 could increase the cell proliferation in the 3D culture.Strangely,we also found more dead cells in the BMSC-collagen sponge constructs that were treated with TGF-β1.Moreover,TGF-β1 promoted more collagen deposition in both the 2D and 3D cultures.CONCLUSION Collagen sponge-based 3D culture with TGF-β1 enhances the responsiveness of the proliferation and tenogenic differentiation of rat BMSCs.     

Hemodynamic simulation of intracranial aneurysm growth with virtual silk stent implantation

Abstract

Longitudinal hemodynamic assessment of intracranial aneurysms (IAs) with endovascular treatment is essential for clinical decision making. We proposed a hemodynamic simulation method to explore the hemodynamic characteristics of a certain growth process of IA with virtual Silk stent implantation. Eight growth point models of IA were created by applying a bidirectional linear growing algorithm on patient 3D rotational angiography images. Simulated experiments showed that the WSS and velocity were significantly decreased with virtual Silk stent implantation at all growth points considered. The simulations revealed the flow pattern and WSS characteristics of IA growth.     

IGF-binding proteins 3 and 4 are regulators of sprouting angiogenesis

Purpose

We have previously identified insulin-like growth factor 2 (IGF2) and insulin-like growth factor 1 receptor (IGF1R) as essential proteins for tip cell maintenance and sprouting angiogenesis. In this study, we aim to identify other IGF family members involved in endothelial sprouting angiogenesis.

Methods

Effects on sprouting were analyzed in human umbilical vein endothelial cells (HUVECs) using the spheroid-based sprouting model, and were quantified as mean number of sprouts per spheroid and average sprout length. RNA silencing technology was used to knockdown gene expression. Recombinant forms of the ligands (IGF1 and IGF2, insulin) and the IGF-binding proteins (IGFBP) 3 and 4 were used to induce excess effects. Effects on the tip cell phenotype were analyzed by measuring the fraction of CD34⁺ tip cells using flow cytometry and immunohistochemistry in a 3D angiogenesis model. Experiments were performed in the presence and absence of serum.

Results

Knockdown of IGF2 inhibited sprouting in HUVECs, in particular when cultured in the absence of serum, suggesting that components in serum influence the signaling of IGF2 in angiogenesis in vitro. We then determined the effects of IGFBP3 and IGFBP4, which are both present in serum, on IGF2-IGF1R signaling in sprouting angiogenesis in the absence of serum: knockdown of IGFBP3 significantly reduced sprouting angiogenesis, whereas knockdown of IGFBP4 resulted in increased sprouting angiogenesis in both flow cytometry analysis and immunohistochemical analysis of the 3D angiogenesis model. Other IGF family members except INSR did not affect IGF2-IGF1R signaling.

Conclusions

Serum components and IGF binding proteins regulate IGF2 effects on sprouting angiogenesis. Whereas IGFBP3 acts as co-factor for IGF2-IGF1R binding, IGFBP4 inhibits IGF2 signaling.

     

Report of the Annual Conference of the Cryptogamic Section, 1948

Background: Trait-based assembly rules are a powerful tool in community ecology, used to explore the pattern and process of community structure (richness and composition).

Aims: A preliminary test for the utility of trait-based assembly rules in explaining cryptogamic epiphyte communities (lichens and bryophytes).

Methods: We sampled epiphytes from three different tree species (aspen, birch and pine), and from trees of contrasting age. The community composition of epiphyte species (taxon analysis) and functional groups (trait analysis) was summarised using multivariate ordination (nonmetric multidimensional scaling, NMDS).

Results: Ordination documented a widely observed pattern in which different tree species have taxonomically different epiphyte communities. However, NMDS sample scores were correlated to tree age in the trait-based analysis, but not in the taxon analysis.

Conclusions: Our results point to the existence of a common pattern in community traits during succession (on trees of different age) when measured for epiphyte communities with contrasting taxonomic composition. This pattern is evidenced by consistent trends in lichen growth form and reproductive strategy (sexual vs. asexual).     

耐镉促生菌株的分离鉴定及对大豆的促生效应

【背景】华南地区镉（Cd）污染严重,与有益微生物共生能够使作物通过直接或间接的机制解除镉毒,提高抗逆性,进而促进生长。耐镉促生菌剂具有广泛的应用前景。【目的】从华南地区受镉污染植株的根内和根际筛选出耐镉且能促进大豆生长的促生菌,以丰富促进田间大豆生产的优异菌种资源。【方法】采用平板划线法从植株的根内或根际分离菌株,通过生理生化特性和16S rRNA基因序列分析对分离菌株进行初步研究,利用盆栽试验探究镉胁迫下菌株对大豆生长的影响,通过测定丙二醛含量和总抗氧化能力探究菌株的耐镉机制。【结果】分离获得4株菌D1、D2、D3和D4,促生特性试验证明4株菌均具有溶磷、产吲哚乙酸和铁载体的能力。经16S rRNA基因序列分析鉴定,D1、D2、D3和D4菌株分别属于不动杆菌属、微小杆菌属、类芽孢杆菌属和普罗威登斯菌属。用这4株菌进行不同镉处理的大豆（巴西10号）盆栽试验,结果表明,4株菌均具有耐镉和促进大豆生长的作用。不添加镉的条件下,大豆接种D4菌株的地上部干重、根部干重和株高分别增加了28%、35%和31%;在添加20mg/kg-CdCl₂·5/2H₂...     

Three-dimensional multicellular cell culture for anti-melanoma drug screening: focus on tumor microenvironment

AbstractThe development of new treatments for malignant melanoma, which has the worst prognosis among skin neoplasms, remains a challenge. The tumor microenvironment aids tumor cells to grow and resist to chemotherapeutic treatment. One way to mimic and study the tumor microenvironment is by using three-dimensional (3D) co-culture models (spheroids). In this study, a melanoma heterospheroid model composed of cancer cells, fibroblasts, and macrophages was produced by liquid-overlay technique using the agarose gel. The size, growth, viability, morphology, cancer stem-like cells population and inflammatory profile of tumor heterospheroids and monospheroids were analyzed to evaluate the influence of stromal cells on these parameters. Furthermore, dacarbazine cytotoxicity was evaluated using spheroids and two-dimensional (2D) melanoma model. After finishing the experiments, it was observed the M2 macrophages induced an anti-inflammatory microenvironment in heterospheroids; fibroblasts cells support the formation of the extracellular matrix, and a higher percentage of melanoma CD271 was observed in this model. Additionally, melanoma spheroids responded differently to the dacarbazine than the 2D melanoma culture as a result of their cellular heterogeneity and 3D structure. The 3D model was shown to be a fast and reliable tool for drug screening, which can mimic the in vivo tumor microenvironment regarding interactions and complexity.Graphic abstract      

Reconstruction of low-resolution molecular structures from simulated atomic force microscopy images

BackgroundAtomic Force Microscopy (AFM) is an experimental technique to study structure-function relationship of biomolecules. AFM provides images of biomolecules at nanometer resolution. High-speed AFM experiments produce a series of images following dynamics of biomolecules. To further understand biomolecular functions, information on three-dimensional (3D) structures is beneficial.MethodWe aim to recover 3D information from an AFM image by computational modeling. The AFM image includes only low-resolution representation of a molecule; therefore we represent the structures by a coarse grained model (Gaussian mixture model). Using Monte-Carlo sampling, candidate models are generated to increase similarity between AFM images simulated from the models and target AFM image.ResultsThe algorithm was tested on two proteins to model their conformational transitions. Using a simulated AFM image as reference, the algorithm can produce a low-resolution 3D model of the target molecule. Effect of molecular orientations captured in AFM images on the 3D modeling performance was also examined and it is shown that similar accuracy can be obtained for many orientations.ConclusionsThe proposed algorithm can generate 3D low-resolution protein models, from which conformational transitions observed in AFM images can be interpreted in more detail.General significanceHigh-speed AFM experiments allow us to directly observe biomolecules in action, which provides insights on biomolecular function through dynamics. However, as only partial structural information can be obtained from AFM data, this new AFM based hybrid modeling method would be useful to retrieve 3D information of the entire biomolecule.     

Source-sink relations affect growth but not the allocation pattern of birch (Betula pendula Roth) seedlings under elevated [CO2]

ABSTRACT

Birch (Betula pendula Roth.) seedlings were kept for two growing seasons under ambient (～350 µmol mol^-1) and elevated (～700 µmol mol^-1) [CO₂]. The present study was designed to examine the effects of [CO₂] and pot size on growth and carbon allocation under conditions of non-limiting water and nutrient supply, in order to separate the effects of source-sink interaction from the effects of nutrient deficiency. The manipulation of the source-sink relations had a strong influence on the growth response to elevated [CO₂]. When the rooting volume was inadequate, it resulted in a source-sink imbalance which constrained growth under elevated [CO₂]. When root exploration was unconstrained, total dry mass was significantly increased (by about 24%) under elevated [CO₂]. However, the allometric relationships in allocation pattern and in morphogenetic development were not affected by either [CO₂] or pot treatments when the saplings were of the same size. Thus, by constraining dry mass production, small sinks affected the magnitude of the growth responses to elevated [CO₂], but did not affect the plant allocation pattern and allometric relationships when nutrient supply was non-limiting. However, by slowing down growth, sink restrictions counteract the speed-up of ontogeny which is the main effect of elevated [CO₂] on tree growth.     

Spatiotemporal effects of Hurricane Ivan on an endemic epiphytic orchid: 10 years of follow-up

ABSTRACT

Background: Hurricanes have a strong influence on the ecological dynamics and structure of tropical forests. Orchid populations are especially vulnerable to these perturbations due to their canopy exposure and lack of underground storage organs and seed banks.

Aims: We evaluated the effects of Hurricane Ivan on the population of the endemic epiphytic orchid Encyclia bocourtii to propose a management strategy.

Methods: Using a pre- and post-hurricane dataset (2003–2013), we assessed the population asymptotic and transient dynamics. We also identified the individual size-stages that maximise population inertia and E. bocourtii’s spatial arrangement relative to phorophytes and other epiphytes.

Results: Hurricane Ivan severely affected the survival and growth of individuals of E. bocourtii, and caused an immediate decline of the population growth rate from λ = 1.05 to λ = 0.32, which was buffered by a population reactivity of ρ₁ = 1.42. Our stochastic model predicted an annual population decrease of 14%. We found an aggregated spatial pattern between E. bocourtii and its host trees, and a random pattern relative to other epiphytes.

Conclusion: Our findings suggest that E. bocourtii is not safe from local extinction. We propose the propagation and reintroduction of reproductive specimens, the relocation of surviving individuals, and the establishment of new plantations of phorophytes.     

Two distinct filopodia populations at the growth cone allow to sense nanotopographical extracellular matrix cues to guide neurite outgrowth

Background

The process of neurite outgrowth is the initial step in producing the neuronal processes that wire the brain. Current models about neurite outgrowth have been derived from classic two-dimensional (2D) cell culture systems, which do not recapitulate the topographical cues that are present in the extracellular matrix (ECM) in vivo. Here, we explore how ECM nanotopography influences neurite outgrowth.

Methodology/Principal Findings

We show that, when the ECM protein laminin is presented on a line pattern with nanometric size features, it leads to orientation of neurite outgrowth along the line pattern. This is also coupled with a robust increase in neurite length. The sensing mechanism that allows neurite orientation occurs through a highly stereotypical growth cone behavior involving two filopodia populations. Non-aligned filopodia on the distal part of the growth cone scan the pattern in a lateral back and forth motion and are highly unstable. Filopodia at the growth cone tip align with the line substrate, are stabilized by an F-actin rich cytoskeleton and enable steady neurite extension. This stabilization event most likely occurs by integration of signals emanating from non-aligned and aligned filopodia which sense different extent of adhesion surface on the line pattern. In contrast, on the 2D substrate only unstable filopodia are observed at the growth cone, leading to frequent neurite collapse events and less efficient outgrowth.

Conclusions/Significance

We propose that a constant crosstalk between both filopodia populations allows stochastic sensing of nanotopographical ECM cues, leading to oriented and steady neurite outgrowth. Our work provides insight in how neuronal growth cones can sense geometric ECM cues. This has not been accessible previously using routine 2D culture systems.     

The temperature‐size rule in Daphnia magna across different genetic lines and ontogenetic stages: Multiple patterns and mechanisms

Ectotherms tend to grow faster, but reach a smaller size when reared under warmer conditions. This temperature‐size rule (TSR) is a widespread phenomenon. Despite the generality of this pattern, no general explanation has been found. We therefore tested the relative importance of two proposed mechanisms for the TSR: (1) a stronger increase in development rate relative to growth rate at higher temperatures, which would cause a smaller size at maturity, and (2) resource limitation placing stronger constraints on growth in large individuals at higher temperatures, which would cause problems with attaining a large size in warm conditions. We raised Daphnia magna at eight temperatures to assess their size at maturity, asymptotic size, and size of their offspring. We used three clonal lines that differed in asymptotic size and growth rate. A resource allocation model was developed and fitted to our empirical data to explore the effect of both mechanisms for the TSR. The genetic lines of D. magna showed different temperature dependence of growth and development rates resulting in different responses for size at maturity. Also, at warm temperatures, growth was constrained in large, but not in small individuals. The resource allocation model could fit these empirical data well. Based on our empirical results and model explorations, the TSR of D. magna at maturity is best explained by a stronger increase in development rate relative to growth rate at high temperature, and the TSR at asymptotic size is best explained by a size‐dependent and temperature‐dependent constraint on growth, although resource limitation could also affect size at maturity. In conclusion, the TSR can take different forms for offspring size, size at maturity, and asymptotic size and each form can arise from its own mechanism, which could be an essential step toward finding a solution to this century‐old puzzle.     

Human Lung Cancer Cells Grown in an Ex Vivo 3D Lung Model Produce Matrix Metalloproteinases Not Produced in 2D Culture

We compared the growth of human lung cancer cells in an ex vivo three-dimensional (3D) lung model and 2D culture to determine which better mimics lung cancer growth in patients. A549 cells were grown in an ex vivo 3D lung model and in 2D culture for 15 days. We measured the size and formation of tumor nodules and counted the cells after 15 days. We also stained the tissue/cells for Ki-67, and Caspase-3. We measured matrix metalloproteinase (MMP) levels in the conditioned media and in blood plasma from patients with adenocarcinoma of the lung. Organized tumor nodules with intact vascular space formed in the ex vivo 3D lung model but not in 2D culture. Proliferation and apoptosis were greater in the ex vivo 3D lung model compared to the 2D culture. After 15 days, there were significantly more cells in the 2D culture than the 3D model. MMP-1, MMP-9, and MMP-10 production were significantly greater in the ex vivo 3D lung model. There was no production of MMP-9 in the 2D culture. The patient samples contained MMP-1, MMP-2, MMP-9, and MMP-10. The human lung cancer cells grown on ex vivo 3D model form perfusable nodules that grow over time. It also produced MMPs that were not produced in 2D culture but seen in human lung cancer patients. The ex vivo 3D lung model may more closely mimic the biology of human lung cancer development than the 2D culture.     

乳腺癌体外骨转移瘤3D模型的建立

目的:构建乳腺癌体外骨转移瘤3D模型。方法:新生CD-1小鼠的颅骨单独孵育为正常组,新生CD-1小鼠的颅骨与MDA-MB-231细胞低氧共孵育四天为模型组。通过扫描电镜(SEM)鉴定骨转移瘤模型,中性红染色法鉴定骨组织中的破骨细胞,硝酸银复染法观察骨的溶解,结晶紫染色法观察骨转移瘤模型中肿瘤细胞的生长。结果:正常组骨表面光滑完整;模型组骨组织表面黏附大量肿瘤细胞,破骨细胞活性增强,产生严重的溶骨,表面出现骨陷窝,骨纤维发生断裂。结论:成功建立了乳腺癌骨转移瘤体外3D模型,该模型能够模拟体内骨转移瘤微环境。     

Conditioned medium derived from 3D tooth germs: A novel cocktail for stem cell priming and early in vivo pulp regeneration

ObjectivesConditioned medium (CM) from 2D cell culture can mitigate the weakened regenerative capacity of the implanted stem cells. However, the capacity of 3D CM to prime dental pulp stem cells (DPSCs) for pulp regeneration and its protein profile are still elusive. We aim to investigate the protein profile of CM derived from 3D tooth germs, and to unveil its potential for DPSCs‐based pulp regeneration.Materials and MethodsWe prepared CM of 3D ex vivo cultured tooth germ organs (3D TGO‐CM) and CM of 2D cultured tooth germ cells (2D TGC‐CM) and applied them to prime DPSCs. Influences on cell behaviours and protein profiles of CMs were compared. In vivo pulp regeneration of CMs‐primed DPSCs was explored using a tooth root fragment model on nude mice.ResultsTGO‐CM enhanced DPSCs proliferation, migration, in vitro mineralization, odontogenic differentiation, and angiogenesis performances. The TGO‐CM group generated superior pulp structures, more odontogenic cells attachment, and enhanced vasculature at 4 weeks post‐surgery, compared with the TGC‐CM group. Secretome analysis revealed that TGO‐CM contained more odontogenic and angiogenic growth factors and fewer pro‐inflammatory cytokines. Mechanisms leading to the differential CM profiles may be attributed to the cytokine–cytokine receptor interaction and PI3K‐Akt signalling pathway.ConclusionsThe unique secretome profile of 3D TGO‐CM made it a successful priming cocktail to enhance DPSCs‐based early pulp regeneration.     

Differential copper binding to alpha-synuclein and its disease-associated mutants affect the aggregation and amyloid formation

BackgroundCopper is an essential trace element required for the proper functioning of various enzymes present in the central nervous system. An imbalance in the copper homeostasis results in the pathology of various neurodegenerative disorders including Parkinson’s Disease. Hence, residue specific interaction of Cu²⁺ to α-Syn along with the familial mutants H50Q and G51D needs to be studied in detail.MethodsWe investigated the residue specific mapping of Cu²⁺ binding sites and binding strength using solution-state NMR and ITC respectively. The aggregation kinetics, secondary structural changes, and morphology of the formed fibrils in the presence and absence of Cu²⁺ were studied using fluorescence, CD, and AFM respectively.ResultsCopper binding to α-Syn takes place at three different sites with a higher affinity for the region 48-53. While one of the sites got abolished in the case of H50Q, the mutant G51D showed a binding pattern similar to WT. The aggregation kinetics of these proteins in the presence of Cu²⁺ showed an enhanced rate of fibril formation with a pronounced effect for G51D.ConclusionCu²⁺ binding results in the destabilization of long-range tertiary interactions in α-Syn leading to the exposure of highly amyloidogenic NAC region which results in the increased rate of fibril formation. Although the residues 48-53 have a stronger affinity for Cu²⁺ in case of WT and G51D, the binding is not responsible for enhancing the rate of fibril formation in case of H50Q.General SignificanceThese findings will help in the better understanding of Cu²⁺ catalyzed aggregation of synucleins.