Mutations in the mitochondrial protein kinase PINK1 are associated with autosomal recessive Parkinson disease (PD). We and other groups have reported that PINK1 activates Parkin E3 ligase activity ...both directly via phosphorylation of Parkin serine 65 (Ser65)—which lies within its ubiquitin‐like domain (Ubl)—and indirectly through phosphorylation of ubiquitin at Ser65. How Ser65‐phosphorylated ubiquitin (ubiquitinPhospho‐Ser65) contributes to Parkin activation is currently unknown. Here, we demonstrate that ubiquitinPhospho‐Ser65 binding to Parkin dramatically increases the rate and stoichiometry of Parkin phosphorylation at Ser65 by PINK1 in vitro. Analysis of the Parkin structure, corroborated by site‐directed mutagenesis, shows that the conserved His302 and Lys151 residues play a critical role in binding of ubiquitinPhospho‐Ser65, thereby promoting Parkin Ser65 phosphorylation and activation of its E3 ligase activity in vitro. Mutation of His302 markedly inhibits Parkin Ser65 phosphorylation at the mitochondria, which is associated with a marked reduction in its E3 ligase activity following mitochondrial depolarisation. We show that the binding of ubiquitinPhospho‐Ser65 to Parkin disrupts the interaction between the Ubl domain and C‐terminal region, thereby increasing the accessibility of Parkin Ser65. Finally, purified Parkin maximally phosphorylated at Ser65 in vitro cannot be further activated by the addition of ubiquitinPhospho‐Ser65. Our results thus suggest that a major role of ubiquitinPhospho‐Ser65 is to promote PINK1‐mediated phosphorylation of Parkin at Ser65, leading to maximal activation of Parkin E3 ligase activity. His302 and Lys151 are likely to line a phospho‐Ser65‐binding pocket on the surface of Parkin that is critical for the ubiquitinPhospho‐Ser65 interaction. This study provides new mechanistic insights into Parkin activation by ubiquitinPhospho‐Ser65, which could aid in the development of Parkin activators that mimic the effect of ubiquitinPhospho‐Ser65.
Synopsis
Binding of phosphorylated ubiquitin to Parkin is shown to induce a conformational change that allows Parkin phosphorylation and maximal activation by PINK1. The study also identifies two residues crucial for the interaction between Parkin and ubiquitinphospho‐Ser65.
Monoubiquitin, as well as diverse ubiquitin chain topologies phosphorylated at serine 65, enhance Parkin phosphorylation by PINK1.
Parkin His302 and Lys151 are required for optimal binding of phospho‐ubiquitin and likely line a phospho‐Ser65‐binding pocket of Parkin.
Parkin phosphorylation and activation at mitochondria is dependent on phospho‐ubiquitin binding.
Binding of phosphorylated ubiquitin to Parkin is shown to induce a conformational change that allows Parkin phosphorylation and maximal activation by PINK1. Two residues crucial for the interaction between Parkin and ubiquitinphospho‐Ser65 are identified.
Hyperexcitability of the central nervous system (CNS) has been suggested to play an important role in the chronic pain experienced by osteoarthritis (OA) patients. A systematic review following ...PRISMA guidelines was performed to evaluate the existing evidence from the literature related to the presence of central sensitization (CS) in patients with OA.Electronic databases PubMed and Web of Science were searched to identify relevant articles using pre‐defined keywords regarding CS and OA. Full‐text clinical reports addressing studies of CS in human adults with chronic complaints due to osteoarthritis were included and screened for methodological quality by two independent reviewers. From the 40 articles that were initially eligible for methodological quality assessment, 36 articles achieved sufficient scores and therefore were discussed. The majority of these studies were case–control studies and addressed OA of the knee joint. Different subjective and objective parameters considered manifestations of CS, which have been previously reported in other chronic pain conditions such as whiplash or rheumatoid arthritis, were established in subjects with OA pain. Overall results suggest that, although peripheral mechanisms are involved in OA pain, hypersensitivity of the CNS plays a significant role in a subgroup of subjects within this population. Although the majority of the literature provides evidence for the presence of CS in chronic OA pain, clinical identification and treatment of CS in OA is still in its infancy, and future studies with good methodological quality are necessary.
Abstract
Industry 4.0 technologies are transforming supply chain management from a linear model to what is known as supply chain 4.0, where operations are integrated, and flows stream in multiple ...directions. The study aims to explore the attributes of Industry 4.0 technologies and their impact to drive sustainability values. The findings were generated after a systematic literature review of 71 articles on various sustainability dimensions and multiple Industry 4.0 technologies. The study provides a review of the most studied technologies in the sustainable supply chain context and categorizes their attributes as well as their relevance to sustainability. The results show that advances in technologies such as Blockchain and Internet of Things have increased the potential for supply chains to reach sustainability values.
The study expands the existing literature and encourages businesses and the scientific community to investigate the power of Industry 4.0 technologies for sustainability.
It presents limitations and directions for future research.
Cities occupy 2% of the earth's surface but their inhabitants consume 75% of the world's energy resources. Under certain conditions, the heat from solar radiation and different urban activities can ...make city temperatures rise in certain areas, simply because of the way in which a city is structured. This effect is known as the urban heat island (UHI). This article provides a review of recent research on the urban heat island as well as of the strategies that can be applied to mitigate its adverse effects. Such strategies can be applied in the project design phase of urban planning and thus directly affect city temperatures on a local scale. The elements analyzed in this paper include green spaces, trees, albedo, pavement surfaces, vegetation, as well as building types and materials. The discussion of this research clearly reflects the impact of urban morphology on local temperatures and how urban design can be modified to reduce energy consumption and CO2 emissions into the atmosphere. This study is useful for professionals who are responsible for decision-making during the design phase of urban planning.
While mean-field approximations, such as the nuclear shell model, provide a good description of many bulk nuclear properties, they fail to capture the important effects of nucleon–nucleon ...correlations such as the short-distance and high-momentum components of the nuclear many-body wave function1. Here, we study these components using the effective pair-based generalized contact formalism2,3 and ab initio quantum Monte Carlo calculations of nuclei from deuteron to 40Ca (refs. 4–6). We observe a universal factorization of the many-body nuclear wave function at short distance into a strongly interacting pair and a weakly interacting residual system. The residual system distribution is consistent with that of an uncorrelated system, showing that short-distance correlation effects are predominantly embedded in two-body correlations. Spin- and isospin-dependent ‘nuclear contact terms’ are extracted in both coordinate and momentum space for different realistic nuclear potentials. The contact coefficient ratio between two different nuclei shows very little dependence on the nuclear interaction model. These findings thus allow extending the application of mean-field approximations to short-range correlated pair formation by showing that the relative abundance of short-range pairs in the nucleus is a long-range (that is, mean field) quantity that is insensitive to the short-distance nature of the nuclear force.Effects of nucleon–nucleon correlations are studied with the generalized contact formalism and ab initio quantum Monte Carlo calculations. For nuclei from deuteron to 40Ca, the many-body nuclear wave function is shown to factorize at short distances.
Extracellular vesicles (EVs) are small membrane-bound vesicles which play an important role in cell-to-cell communication. Their molecular cargo analysis is presented as a new source for biomarker ...detection, and it might provide an alternative to traditional solid biopsies. However, the most effective approach for EV isolation is not yet well established.
Here, we study the efficiency of the most common EV isolation methods-ultracentrifugation, Polyethlyene glycol and two commercial kits, Exoquick
and PureExo
. We isolated circulating EVs from the bloodstream of healthy donors, characterized the size and yield of EVs and analyzed their protein profiles and concentration. Moreover, we have used for the first time Digital-PCR to identify and detect specific gDNA sequences, which has several implications for diagnostic and monitoring many types of diseases.
Our findings present Polyethylene glycol precipitation as the most feasible and less cost-consuming EV isolation technique.
During recent years, advances in drone technologies have made them applicable in various fields of industry, and their popularity continues to grow. In this paper, the academic contributions on ...drones routing problems are analyzed between 2005 and 2019 to identify the main characteristics of these types of problems, as well as the research trends and recent improvements. The literature is classified according to the objectives optimized, solution methods, applications, constraints, and whether they use a complementary vehicle or not. Finally, a discussion for trends and future research is presented.
The PARK2 gene is mutated in 50% of autosomal recessive juvenile parkinsonism (ARJP) cases. It encodes parkin, an E3 ubiquitin ligase of the RBR family. Parkin exists in an autoinhibited state that ...is activated by phosphorylation of its N‐terminal ubiquitin‐like (Ubl) domain and binding of phosphoubiquitin. We describe the 1.8 Å crystal structure of human parkin in its fully inhibited state and identify the key interfaces to maintain parkin inhibition. We identify the phosphoubiquitin‐binding interface, provide a model for the phosphoubiquitin–parkin complex and show how phosphorylation of the Ubl domain primes parkin for optimal phosphoubiquitin binding. Furthermore, we demonstrate that the addition of phosphoubiquitin leads to displacement of the Ubl domain through loss of structure, unveiling a ubiquitin‐binding site used by the E2~Ub conjugate, thus leading to active parkin. We find the role of the Ubl domain is to prevent parkin activity in the absence of the phosphorylation signals, and propose a model for parkin inhibition, optimization for phosphoubiquitin recruitment, release of inhibition by the Ubl domain and engagement with an E2~Ub conjugate. Taken together, this model provides a mechanistic framework for activating parkin.
Synopsis
Three dimensional structures of the E3 ubiquitin ligase parkin in its auto‐inhibited, optimized and activated forms show how its ubiquitin‐like domain (Ubl) is the key controller of enzyme function, being allosterically displaced through phospho‐ubiquitin recruitment to unveil a masked E2~Ub binding pocket required for full ubiquitination activity.
The Ubl domain keeps parkin kept inactive in the absence of phosphorylation signals.
A hinge in the centre of parkin optimizes a phospho‐ubiquitin binding patch at the RING0‐RING1 interface.
Phospho‐ubiquitin binding along a unique helix in the RING1 domain releases the inhibitory Ubl domain.
Ubl domain displacement reveals a ubiquitin‐binding surface that enhances E2~Ub engagement.
Structural details show how parkin function can be altered by mutations in early‐onset Parkinson's disease.
Three‐dimensional structures of parkin in various states reveal key control by its ubiquitin‐like domain (Ubl), which is allosterically displaced through phospho‐ubiquitin recruitment.
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