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.
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.
Mutations in the park2 gene, encoding the RING-inBetweenRING-RING E3 ubiquitin ligase parkin, cause 50% of autosomal recessive juvenile Parkinsonism cases. More than 70 known pathogenic mutations ...occur throughout parkin, many of which cluster in the inhibitory amino-terminal ubiquitin-like domain, and the carboxy-terminal RING2 domain that is indispensable for ubiquitin transfer. A structural rationale showing how autosomal recessive juvenile Parkinsonism mutations alter parkin function is still lacking. Here we show that the structure of parkin RING2 is distinct from canonical RING E3 ligases and lacks key elements required for E2-conjugating enzyme recruitment. Several pathogenic mutations in RING2 alter the environment of a single surface-exposed catalytic cysteine to inhibit ubiquitination. Native parkin adopts a globular inhibited conformation in solution facilitated by the association of the ubiquitin-like domain with the RING-inBetweenRING-RING C-terminus. Autosomal recessive juvenile Parkinsonism mutations disrupt this conformation. Finally, parkin autoubiquitinates only in cis, providing a molecular explanation for the recessive nature of autosomal recessive juvenile Parkinsonism.
ABSTRACT
An analytical study was conducted to assess the long‐term influence, role, and impacts of El Niño‐Southern Oscillation (ENSO) on Puerto Rico's precipitation patterns and significant moisture ...deficits (droughts). Detection and attribution was addressed by evaluating local rainfall measures and ENSO‐related data to (1) detect ENSO signals and patterns, (2) quantify the magnitude of any impacts, and (3) determine if ENSO may be an important factor for local prediction of future droughts. Data were evaluated at different time periods and two spatial scales (island‐wide and internal climate regions of Puerto Rico). Although a signal was detected, it was weak, in both directions, varied regionally, and has inconsequential impacts. No evidence was found for a major control by ENSO over local monthly, seasonal, and yearly rainfall for any climate regions on the island. These results indicate that ENSO is not a main factor causing droughts in Puerto Rico for the study period and thus should not be a factor in predicting the potential for local dry periods or large precipitation deficits in the future. Any presumed teleconnections between Puerto Rico's dry periods and ENSO are not based on current climatological evidence. Thus, local drought prediction efforts should be focused on finding major causes of local rainfall variation other than ENSO.
ENSO's signal was detected in Puerto Rico using different methods but actual impacts on rainfall for the past century are minimal to none at any time and spatial scales.
PDF
