Intrinsic Enzymatic Activity Research Articles

Doxorubicin as a Drug Repurposing for Disruption of α-Chymotrypsinogen-A Aggregates.

Protein conformation is affected by interaction of several small molecules resulting either stabilization or disruption depending on the nature of the molecules. In our earlier communication, Hg2+ was known to disrupt the native structure of α-Cgn A leading to aggregation (Ansari, N.K., Rais, A. & Naeem, A. Methotrexate for Drug Repurposing as an Anti-Aggregatory Agent to Mercuric Treated α-Chymotrypsinogen-A. Protein J (2024). https://doi.org/10.1007/s10930-024-10187-z ). Accumulation of β-rich aggregates in the living system is found to be linked with copious number of disorders. Here, we have investigated the effect of varying concentration of doxorubicin (DOX) i.e. 0-100 µM on the preformed aggregates of α-Cgn A upon incubation with 120 µM Hg2+. The decrease in the intrinsic fluorescence and enzyme activity with respect to increase in the Hg2+ concentration substantiate the formation of aggregates. The DOX showed the dose dependent decrease in the ThT fluorescence, turbidity and RLS measurements endorsing the dissolution of aggregates which were consistent with red shift in ANS, confirming the breakdown of aggregates. The α-Cgn A has 30% α-helical content which decreases to 3% in presence of Hg2+. DOX increased the α-helicity to 28% confirming its anti-aggregatory potential. The SEM validates the formation of aggregates with Hg2+ and their dissolution upon incubation with the DOX. Hemolysis assay checked the cytotoxicity of α-Cgn A aggregates. Docking revealed that the DOX interacted Lys203, Cys201, Cys136, Ser159, Leu10, Trp207, Val137 and Thr134 of α-Cgn A through hydrophobic interactions and Gly133, Thr135 and Lys202 forms hydrogen bonds.

Abstract PR007: Identifying targeted therapies for muscle invasive bladder cancer via STAG2 expression

Abstract High stromal antigen 2 (STAG2) protein expression is significantly associated with shorter overall and progression free survival of muscle invasive bladder cancer (MIBC) patients. Unfortunately, STAG2 itself is difficult to target as it functions as a transcriptional regulator and has no intrinsic enzymatic activity. Therefore, it becomes essential to identify downstream therapeutic vulnerabilities based on STAG2 expression. In this study, we accomplish this by employing a drug screen in isogenic STAG2 wild-type (WT) and knock-out (KO) MIBC cell lines, analyzing drug sensitivity of DepMap bladder cancer cell lines, and performing in vitro dose-response experiments based on our drug screen findings. To delineate changes in drug sensitivity due specifically to altered STAG2 expression, we used CRISPR-Cas9 to knock out STAG2 expression in T24 MIBC cells and generated two clonal STAG2 KO cell lines to compare to control. Then, we performed a drug screen with 312 drugs targeting oncogenic signaling pathways. We treated cells with drugs at 534 nM and read viability after 72 hours. To expand our drug screen findings to a larger group of cell lines and drugs, we further queried the DepMap database which includes drug sensitivity data for 4,868 drugs and approximately 2,000 cancer cell lines. We selected drug screened bladder cancer cell lines from DepMap and grouped them into ‘STAG2-high’ or ‘STAG2-low’ cohorts based on STAG2 mRNA expression. We then compared these two groups to identify if either group was more sensitive to drugs in the DepMap database. From these drug screen and DepMap analyses, we identified TAK-733 (MEKi), Berzosertib (ATRi), Prexasertib (CHKi), PI-103 (PI3Ki), and Rigosertib (PLKi) as candidate drugs that are differentially effective based on STAG2 expression level. We performed validation studies with these candidate drugs by treating matched STAG2 WT and KO T24 cells with each drug at concentrations ranging from 0.001 to 100 µM, then computationally modeled EC50 values in R. Cells with STAG2 KO were more sensitive to Berzosertib (T24 Control EC50: 1.78 µM, T24 KO EC50s: 1.03, 0.77 µM) compared to cells with WT STAG2 expression. Interestingly, cells with WT STAG2 expression were more sensitive to PI-103 (T24 Control EC50: 0.33 µM, T24 KO EC50s: 0.97, 1.41 µM) compared to cells with STAG2 KO. This suggests that STAG2 expressing cells may have a unique dependency on PI3K signaling, which has previously been reported in sarcoma cell lines. These findings lay the foundation for further investigation of Berzosertib as a targeted therapy for MIBC patients with mutated or low STAG2, and PI-103 for MIBC patients with high STAG2 who currently suffer worse clinical outcomes. Citation Format: Sarah R. Athans, Henry Withers, Zara Kazmierczak, Katerina Gurova, Anna Woloszynska. Identifying targeted therapies for muscle invasive bladder cancer via STAG2 expression [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2024 May 17-20; Charlotte, NC. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(10_Suppl):Abstract nr PR007.

The mechanism of intrinsic peroxidase (POD)-like activity of attapulgite.

Natural attapulgite (ATP) is a promising substitute for existing artificial nanozymes due to its intrinsic enzymatic activity. However, the active center of ATP's inherent enzymatic activity has not yet been revealed, which limits its further design and activity optimization. Studying the active center of mineral materials can be extremely challenging due to their complexity. Here, we demonstrated that Fe is the primary element in ATP responsible for peroxidase (POD)-like activity through theoretical speculation and experimental verification. More importantly, we found that the ratio of Fe2+/Fe3+ is responsible for the district POD-like activity of ATP from different regions with the same Fe content. Additionally, three facile strategies, including grinding, heat treatment, and acid treatment, were demonstrated to increase the relative Fe content and thus optimize the POD-like activity of ATP. Finally, ATP was used to detect the concentration of H2O2, enabling the detection of low concentrations (0.11-1.76mM) of H2O2. This study serves as a novel reference for the future design and performance optimization of nanozymes that are based on ATP and clay minerals.

Comprehensive Review of the Potential of Sorghum in Malt and Beer Production

Sorghum has a relatively lower price compared to other brewing cereals, and consequently has potential as an alternative substrate for lager beer brewing and especially represents an excellent option for gluten-free beers. Attributes such as extracts, proteins, total nitrogen and free amino nitrogen content of sorghum malt and the wort obtained from mashes indicate that sorghum is potentially an alternative substrate for conventional beer brewing in the tropics. In alcohol industry, some distillers prefer sorghum varieties with high starch and less protein contents. The variation in biochemical compositions among sorghum varieties affects the optimal malting conditions and characteristics of the grains. The degree of steeping and temperature, and germination capacity, kilning temperature and the intrinsic enzyme activity determine sorghum grain malt quality. The high degree of gelatinization temperature, poor saccarification, lower fermentability and filterability of sorghum malt could generally affects the acceptability of sorghum beer by the consumer than a beer brewed from malt barley.

Development of tumor marker detection and tumor treatment based on silver nanozymes

Nanomaterials with intrinsic enzyme activity (nanozymes) have been widely used as artificial enzymes in biomedicine, especially in tumor diagnosis and treatment. Herein, a kind of water-soluble silver nanozyme (Ag@GQD) was developed with a simple method, which showed two enzymatic activities (peroxidase and oxidase-like activity). Based on the excellent catalytic performance of oxidase-like activity, a simple and highly sensitive colorimetric detection strategy for glutathione (GSH) was established. What’s more, after absorbing DOX, Ag/DOX@GQD could realize the combined treatment of reactive oxygen species and chemotherapy. In conclusion, this study provided evidences that Ag@GQD is effective in tumor diagnosis and treatment, which provided new ideas for exploring nanozymes based on the integration of diagnosis and treatment.

Organ-specific biotransformation in salmonids: Insight into intrinsic enzyme activity and biotransformation of three micropollutants

Aquatic ecosystems continue to be threatened by chemical pollution. To what extent organisms are able to cope with chemical exposure depends on their ability to display mechanisms of defense across different organs. Among these mechanisms, biotransformation processes represent key physiological responses that facilitate detoxification and reduce the bioaccumulation potential of chemicals. Biotransformation does not only depend on the ability of different organs to display biotransformation enzymes but also on the affinity of chemicals towards these enzymes. In the present study, we explored the ability of different organs and of two freshwater fish to support biotransformation processes through the determination of in vitro phase I and II biotransformation enzyme activity, and their role in supporting intrinsic clearance and the formation of biotransformation products. Three environmentally relevant pollutants were evaluated: the polycyclic aromatic hydrocarbon (PAH) pyrene (as recommended by the OECD 319b test guideline), the fungicide azoxystrobin, and the pharmaceutical propranolol. Comparative studies using S9 sub-cellular fractions derived from the liver, intestine, gills, and brain of brown trout (Salmo trutta) and rainbow trout (Oncorhynchus mykiss) revealed significant phase I and II enzyme activity in all organs. However, organ- and species-specific differences were found. In brown trout, significant extrahepatic biotransformation was observed for pyrene but not for azoxystrobin and propranolol. In rainbow trout, the brain appeared to biotransform azoxystrobin. In this same species, propranolol appeared to be biotransformed by the intestine and gills. Biotransformation products could be detected only from hepatic biotransformation, and their profiles and formation rates displayed species-specific patterns and occurred at different magnitudes. Altogether, our findings further contribute to the current understanding of organ-specific biotransformation capacity, beyond the expression and activity of enzymes, and its dependence on specific enzyme-chemical interactions to support mechanisms of defense against exposure.

A carbon-based bifunctional heterogeneous enzyme: toward sustainable pollution control.

We present a study on an immobilized functional enzyme (IFE), a novel biomaterial with exceptional sustainability in enzyme utility, widely employed across various fields worldwide. However, conventional carriers are prone to eroding the active functional domain of the IFE, thereby weakening its intrinsic enzyme activity. Consequently, there is a burgeoning interest in developing next-generation IFEs. In this study, we engineered a carbon-based bifunctional heterogeneous enzyme (MIP-AMWCNTs@lipase) for the intelligent recognition of di(2-ethylhexyl)phthalate (DEHP), a common plasticizer. The heterogeneous enzyme contains a bifunctional structural domain that both enriches and degrades DEHP. We investigated its dual-response performance for the enrichment and specific removal of DEHP. The imprinting factor of the carrier for DEHP was 3.4, demonstrating selectivity for DEHP. The removal rate reached up to 94.2% over a short period. The heterogeneous enzyme exhibited robust activity, catalytic efficiency, and excellent stability under harsh environmental conditions, retaining 77.7% of its initial lipase activity after 7 cycles. Furthermore, we proposed a stepwise heterogeneous enzyme reaction kinetic model based on the Michaelis-Menten equation to enhance our understanding of enzyme reaction kinetics. Our study employs a dual-effect recognition strategy of molecular blotting and enzyme immobilization to establish a method for the removal of organic pollutants. These findings hold significant implications for the fields of biomaterials and environmental science.

Research progress of new multifunctional bone cement in bone tumor therapy

The research progress of new multifunctional bone cement in bone tumor therapy in recent years was reviewed, in order to provide help for the future research of anti-tumor bone cement. The related literature on the treatment of bone tumors with new multifunctional bone cement at home and abroad in recent years was extensively reviewed and summarized. The new multifunctional bone cements include those with the functions of photothermotherapy, magnetic thermotherapy, chemoradiotherapy, and antibacterial after operation, which are discussed from the aspects of anti-tumor, drug controlled release, and cytotoxicity. Controlled drug release has been achieved in multifunctional bone cements by adjusting heat and pH or incorporating particles such as chitosan oligosaccharides and γ-cyclodextrin. At present, multifunctional bone cement with hyperthermia, radiotherapy, and chemotherapy has effectively inhibited the local recurrence and distant metastasis of bone tumors. Broadening the application of bone cement for photothermal and magnetic thermal therapy to deeper bone tumors, investigating more precise controlled release of drug-loaded bone cement, and introducing nanoparticles with both thermal conversion and intrinsic enzymatic activities into bone cement for synergistic anti-tumor therapy are promising research directions. The new multifunctional bone cement inhibits bone tumor cells, promotes new bone formation in bone defects, and prevents incision infection after tumor resection. Certain progress has been made in anti-tumor, antibacterial, drug-controlled release, and reduction of cytotoxicity. Expanding the deeper application range of the new multifunctional bone cement, verifying the safety in clinical application, and focusing on the individualized treatment of the new multifunctional bone cement are the problems that need to be solved in the future.

Cell cycle and enzymatic activity alterations induced by ROS production in human neuroblastoma cells SH-SY5Y exposed to Fumonisin B1, Ochratoxin A and their combination

The presence of mycotoxins such as Fumonisin B1(FB1) and Ochratoxin A (OTA) in food and feed has become a threat to human and animal health since they can produce several afflictions. Different mechanisms of action by which they exercise their cytotoxic activity have been attributed to them, including the production of reactive oxygen species (ROS). For this reason, a measurement of the production of ROS species, and an evaluation of the intrinsic cell enzymatic antioxidant activity, including glutathione peroxidase (GPx), glutathione transferase (GTS), and catalase (CAT) together with a cytotoxicity and cell cycle assay have been performed in undifferentiated SH-SY5Y cells exposed to FB1, OTA and [FB1 + OTA] after 24 h and 48 h. FB1 and OTA. Monitoring of intracellular ROS production was carried out by the H2-DCFDA probe; while spectrometry analysis of absorbances was used for measuring GPx, GST and CAT activity. Finally, cell proliferation and cell cycle distribution were studied by flow cytometry. When cells were treated with OTA, an increase in GPx and GST activity was observed compared to FB1 and [FB1 + OTA]; conversely, a decrease in CAT activity was observed when cells were exposed to OTA coinciding with the results observed for ROS measurement. Regarding the cell cycle, when cells were exposed to OTA, a decrease in G0/G1 was detected, revealing an arrest of cell division for SH-SY5Y cells at the concentrations studied.

Dihydromyricetin confers cerebroprotection against subarachnoid hemorrhage via the Nrf2-dependent Prx2 signaling cascade

BackgroundSeveral clinical and experimental studies have shown that therapeutic strategies targeting oxidative damage are beneficial for subarachnoid hemorrhage (SAH). A brain-permeable flavonoid, dihydromyricetin (DHM), can modulate redox/oxidative stress and has cerebroprotective effects in several neurological disorders. The effects of DHM on post-SAH early brain injury (EBI) and the underlying mechanism have yet to be clarified. PurposeThis work investigated a potential role for DHM in SAH, together with the underlying mechanisms. MethodsCerebroprotection by DHM was studied using a SAH rat model and primary cortical neurons. Atorvastatin (Ato) was a positive control drug in this investigation. The effects of DHM on behavior after SAH were evaluated by performing the neurological rotarod and Morris water maze tests, as well as by examining its effects on brain morphology and on the molecular and functional phenotypes of primary cortical neurons using dichlorodihydrofluorescein diacetate (DCFH-DA), immunofluorescent staining, biochemical analysis, and Western blot. ResultsDHM was found to significantly reduce the amount of reactive oxygen species (ROS), suppress mitochondrial disruption, and increase intrinsic antioxidant enzymatic activity following SAH. DHM also significantly reduced neuronal apoptosis in SAH rats and improved short- and long-term neurological functions. DHM induced significant increases in peroxiredoxin 2 (Prx2) and nuclear factor erythroid 2-related factor 2 (Nrf2) expression, while decreasing phosphorylation of p38 and apoptotic signal-regulated kinase 1 (ASK1). In contrast, reduction of Prx2 expression using small interfering ribonucleic acid or by inhibiting Nrf2 with ML385 attenuated the neuroprotective effect of DHM against SAH. Moreover, DHM dose-dependently inhibited oxidative damage, decreased neuronal apoptosis, and increased the viability of primary cultured neurons in vitro. These positive effects were associated with Nrf2 activation and stimulation of Prx2 signaling, whereas ML385 attenuated the beneficial effects. ConclusionThese results reveal that DHM protects against SAH primarily by modulating the Prx2 signaling cascade through the Nrf2-dependent pathway. Hence, DHM could be a valuable therapeutic candidate for SAH treatment.

Differential roles of CTP synthetases CTPS1 and CTPS2 in cell proliferation.

The CTP nucleotide is a key precursor of nucleic acids metabolism essential for DNA replication. De novo CTP production relies on CTP synthetases 1 and 2 (CTPS1 and CTPS2) that catalyze the conversion of UTP into CTP. CTP synthetase activity is high in proliferating cells including cancer cells; however, the respective roles of CTPS1 and CTPS2 in cell proliferation are not known. By inactivation of CTPS1 and/or CTPS2 and complementation experiments, we showed that both CTPS1 and CTPS2 are differentially required for cell proliferation. CTPS1 was more efficient in promoting proliferation than CTPS2, in association with a higher intrinsic enzymatic activity that was more resistant to inhibition by 3-deaza-uridine, an UTP analog. The contribution of CTPS2 to cell proliferation was modest when CTPS1 was expressed but essential in absence of CTPS1. Public databases analysis of more than 1,000 inactivated cancer cell lines for CTPS1 or CTPS2 confirmed that cell growth is highly dependent of CTPS1 but less or not of CTPS2. Therefore, our results demonstrate that CTPS1 is the main contributor to cell proliferation.

Identification of WD-Repeat Protein 72 as a Novel Prognostic Biomarker in Non-Small-Cell Lung Cancer.

WD-repeat protein 72(WDR72; OMIM∗613214), a scaffolding protein lacking intrinsic enzymatic activity, produces numerous β-propeller blade formations, serves as a binding platform to assemble protein complexes and is critical for cell growth, differentiation, adhesion, and migration. Despite evidence supporting a basic role of WDR72 in the tumorigenesis of particular cancers, the value of WDR72 in non-small-cell lung cancer (NSCLC), the tumor with the highest mortality rate globally, is undocumented. We investigated the prognostic value of WDR72 in NSCLC and studied its potential immune function and its correlation with ferroptosis. According to The Cancer Genome Atlas, Cancer Cell Line Encyclopedia, Genotype-Tissue Expression, and Gene Set Cancer Analysis, we used multiple bioinformatic strategies to investigate the possible oncogenic role of WDR72, analyze WDR72 and prognosis, and immune cell infiltration in different tumors correlation. WDR72 exhibited a high expression in NSCLC and a positive association with prognosis. WDR72 expression was related to immune cell infiltration and tumor immune microenvironment in NSCLC. Finally, we validated WDR72 in human NSCLC; it has a predictive value in NSCLC related to its function in tumor progression and immunity. The significance of our study is that WDR72 can be used as a potential indicator of lung cancer prognosis. Helping physicians more accurately predict patient survival and risk of disease progression.

Biogenic metallic nanoparticles as enzyme mimicking agents.

The use of biological systems such as plants, bacteria, and fungi for the synthesis of nanomaterials has emerged to fill the gap in the development of sustainable methods that are non-toxic, pollution-free, environmentally friendly, and economical for synthesizing nanomaterials with potential in biomedicine, biotechnology, environmental science, and engineering. Current research focuses on understanding the characteristics of biogenic nanoparticles as these will form the basis for the biosynthesis of nanoparticles with multiple functions due to the physicochemical properties they possess. This review briefly describes the intrinsic enzymatic mimetic activity of biogenic metallic nanoparticles, the cytotoxic effects of nanoparticles due to their physicochemical properties and the use of capping agents, molecules acting as reducing and stability agents and which aid to alleviate toxicity. The review also summarizes recent green synthetic strategies for metallic nanoparticles.

Mitochondrial-nuclear communication by FKBP51 shuttling.

The HSP90-binding immunophilin FKBP51 is a soluble protein that shows high homology and structural similarity with FKBP52. Both immunophilins are functionally divergent and often show antagonistic actions. They were first described in steroid receptor complexes, their exchange in the complex being the earliest known event in steroid receptor activation upon ligand binding. In addition to steroid-related events, several pleiotropic actions of FKBP51 have emerged during the last years, ranging from cell differentiation and apoptosis to metabolic and psychiatric disorders. On the other hand, mitochondria play vital cellular roles in maintaining energy homeostasis, responding to stress conditions, and affecting cell cycle regulation, calcium signaling, redox homeostasis, and so forth. This is achieved by proteins that are encoded in both the nuclear genome and mitochondrial genes. This implies active nuclear-mitochondrial communication to maintain cell homeostasis. Such communication involves factors that regulate nuclear and mitochondrial gene expression affecting the synthesis and recruitment of mitochondrial and nonmitochondrial proteins, and/or changes in the functional state of the mitochondria itself, which enable mitochondria to recover from stress. FKBP51 has emerged as a serious candidate to participate in these regulatory roles since it has been unexpectedly found in mitochondria showing antiapoptotic effects. Such localization involves the tetratricopeptiderepeats domains of the immunophilin and not its intrinsic enzymatic activity of peptidylprolyl-isomerase. Importantly, FKBP51 abandons the mitochondria and accumulates in the nucleus upon cell differentiation or during the onset of stress. Nuclear FKBP51 enhances the enzymatic activity of telomerase. The mitochondrial-nuclear trafficking is reversible, and certain situations such as viral infections promote the opposite trafficking, that is,FKBP51 abandons the nucleus and accumulates in mitochondria. In this article, we review the latest findings related to the mitochondrial-nuclear communication mediated by FKBP51 and speculate about the possible implications of this phenomenon.

Carbon Quantum Dots: Synthesis, Structure, Properties, and Catalytic Applications for Organic Synthesis

Carbon quantum dots (CQDs), also known as carbon dots (CDs), are novel zero-dimensional fluorescent carbon-based nanomaterials. CQDs have attracted enormous attention around the world because of their excellent optical properties as well as water solubility, biocompatibility, low toxicity, eco-friendliness, and simple synthesis routes. CQDs have numerous applications in bioimaging, biosensing, chemical sensing, nanomedicine, solar cells, drug delivery, and light-emitting diodes. In this review paper, the structure of CQDs, their physical and chemical properties, their synthesis approach, and their application as a catalyst in the synthesis of multisubstituted 4H pyran, in azide-alkyne cycloadditions, in the degradation of levofloxacin, in the selective oxidation of alcohols to aldehydes, in the removal of Rhodamine B, as H-bond catalysis in Aldol condensations, in cyclohexane oxidation, in intrinsic peroxidase-mimetic enzyme activity, in the selective oxidation of amines and alcohols, and in the ring opening of epoxides are discussed. Finally, we also discuss the future challenges in this research field. We hope this review paper will open a new channel for the application of CQDs as a catalyst in organic synthesis.

Peroxidase-like activity of silver nanowires and its application for colorimetric detection of the antibiotic chloramphenicol

Different nanomaterials possessing localized surface plasmon resonance (LSPR) such as Ag, Au, Cu, Ti are considered as valuable tools in biomedicine and biotechnology, especially as signal enhancers in different immunoassays. In particular, silver nanoparticles as one of the most effective plasmonic metamaterials are widely used for the development of analytical detection methods, where nanoparticles either act as labels or exhibit intrinsic enzymatic activity (e.g., peroxidase or catalase-like). Silver nanowires (AgNWs) synthesized through polyol synthesis are stable nanomaterials with a highly reactive surface. Here we show that AgNWs possess intrinsic peroxidase-like activity with three commonly used substrates – 3,3’,5,5’-tetramethybenzidine (TMB), o-phenylenediamine (OPD), or diaminobenzidine (DAB) both in precipitating and non-precipitating modes. Moreover, we showed that AgNWs together with OPD can serve as a robust hydrogen peroxide sensor and pH-sensor: AgNW peroxidase-like activity increases linearly with pH. The capabilities of AgNWs as enzyme-like labels were demonstrated in competitive enzyme-linked immunoassay with chloramphenicol (CAP) biosensing with the 0.36 µM (0.15 µg/mL) detection limit. Moreover, the LSPR property of AgNW allows using them as optical labels for the visualization of antibody-hapten interaction – the fact was confirmed with CAP detection in the lateral flow test within the 0.12 ÷ 88 µM working concentration range. Taking into account the unique stability of AgNWs (AgNW were stable for more than two years at room temperature in water without any preservatives and light protection), AgNWs should be considered as robust, convenient, and reliable optical labels for the development of different commercial assays for the wide spectrum of biological applications.

Natural variants of human SARM1 cause both intrinsic and dominant loss-of-function influencing axon survival

SARM1 is a central executioner of programmed axon death, and this role requires intrinsic NAD(P)ase or related enzyme activity. A complete absence of SARM1 robustly blocks axon degeneration in mice, but even a partial depletion confers meaningful protection. Since axon loss contributes substantially to the onset and progression of multiple neurodegenerative disorders, lower inherent SARM1 activity is expected to reduce disease susceptibility in some situations. We, therefore, investigated whether there are naturally occurring SARM1 alleles within the human population that encode SARM1 variants with loss-of-function. Out of the 18 natural SARM1 coding variants we selected as candidates, we found that 10 display loss-of-function in three complimentary assays: they fail to robustly deplete NAD in transfected HEK 293T cells; they lack constitutive and NMN-induced NADase activity; and they fail to promote axon degeneration in primary neuronal cultures. Two of these variants are also able to block axon degeneration in primary culture neurons in the presence of endogenous, wild-type SARM1, indicative of dominant loss-of-function. These results demonstrate that SARM1 loss-of-function variants occur naturally in the human population, and we propose that carriers of these alleles will have different degrees of reduced susceptibility to various neurological conditions.

Self-assembled artificial enzyme from hybridized porous organic cages and iron oxide nanocrystals

HypothesisAlthough it is well-accepted that iron oxide nanoparticles are considered as artificial enzymes when their surface is hydrophilic, the enzyme-like properties of iron oxide nanoparticles with hydrophobic surface coating is unexplored. This work demonstrates that hydrophobic iron oxide nanocrystals coated with a layer of oleic acid could serve as artificial enzymes when their surface is covered by a layer of ionic surfactant. Furthermore, the co-assembly of iron oxide nanocrystals and porous organic cages could modulate their enzyme-like activities. ExperimentsCo-assembly of iron oxide (Fe3O4) nanocrystals with different size and porous organic cages (POCs) was performed by an emulsion-confined strategy to achieve hybridized Fe3O4/POCs co-assemblies. The peroxidase-mimic activity of these co-assemblies were assessed in the presence of 3, 3′, 5, 5′-Tetramethylbenzidine (TMB) and hydrogen peroxide. Finally, these co-assemblies were applied as sensors to detect glucose and hydrogen peroxide. FindingsCo-assembly of Fe3O4 nanocrystals and POCs resulted in the self-assembly of Fe3O4 nanoparticles into two-dimensional nanoparticle superlattices on the eight (111) facets of the octahedral POCs colloidal crystals. The unique oil-in-water (O/W) emulsion confined assembly method switches the Fe3O4 nanoparticles and POC crystals from hydrophobic to hydrophilic because of the strong hydrophobic interactions. Importantly, these co-assemblies dispersed in water showed strong peroxidase-mimic activity in water despite that their surface is covered by a bilayer of aliphatic chains. Furthermore, the intrinsic enzymatic activity of the co-assemblies is highly dependent on the size of the nanocrystals, and a higher catalytic activity is achieved from a larger sized Fe3O4 nanocrystal.

Coreceptor functions of cell surface heparan sulfate proteoglycans.

Receptor-ligand interactions play an important role in many biological processes by triggering specific cellular responses. These interactions are frequently regulated by coreceptors that facilitate, alter, or inhibit signaling. Coreceptors work in parallel with other specific and accessory molecules to coordinate receptor-ligand interactions. Cell surface heparan sulfate proteoglycans (HSPGs) function as unique coreceptors because they can bind to many ligands and receptors through their HS and core protein motifs. Cell surface HSPGs are typically expressed in abundance of the signaling receptors and, thus, are capable of mediating the initial binding of ligands to the cell surface. HSPG coreceptors do not possess kinase domains or intrinsic enzyme activities and, for the most part, binding to cell surface HSPGs does not directly stimulate intracellular signaling. Because of these features, cell surface HSPGs primarily function as coreceptors for many receptor-ligand interactions. Given that cell surface HSPGs are widely conserved, they likely serve fundamental functions to preserve basic physiological processes. Indeed, cell surface HSPGs can support specific cellular interactions with growth factors, morphogens, chemokines, extracellular matrix (ECM) components, and microbial pathogens and their secreted virulence factors. Through these interactions, HSPG coreceptors regulate cell adhesion, proliferation, migration, and differentiation, and impact the onset, progression, and outcome of pathophysiological processes, such as development, tissue repair, inflammation, infection, and tumorigenesis. This review seeks to provide an overview of the various mechanisms of how cell surface HSPGs function as coreceptors.

Role of Microbiota-Gut-Brain Axis in Regulating Dopaminergic Signaling.

Dopamine is a neurotransmitter that plays a critical role both peripherally and centrally in vital functions such as cognition, reward, satiety, voluntary motor movements, pleasure, and motivation. Optimal dopamine bioavailability is essential for normal brain functioning and protection against the development of neurological diseases. Emerging evidence shows that gut microbiota have significant roles in maintaining adequate concentrations of dopamine via intricate, bidirectional communication known as the microbiota-gut-brain axis. The vagus nerve, immune system, hypothalamus–pituitary–adrenal axis, and microbial metabolites serve as important mediators of the reciprocal microbiota-gut-brain signaling. Furthermore, gut microbiota contain intrinsic enzymatic activity that is highly involved in dopamine metabolism, facilitating dopamine synthesis as well as its metabolite breakdown. This review examines the relationship between key genera of gut microbiota such as Prevotella, Bacteroides, Lactobacillus, Bifidobacterium, Clostridium, Enterococcus, and Ruminococcus and their effects on dopamine. The effects of gut dysbiosis on dopamine bioavailability and the subsequent impact on dopamine-related pathological conditions such as Parkinson’s disease are also discussed. Understanding the role of gut microbiota in modulating dopamine activity and bioavailability both in the periphery and in the central nervous system can help identify new therapeutic targets as well as optimize available methods to prevent, delay, or restore dopaminergic deficits in neurologic and metabolic disorders.