Johansenjarvis5806

Orientin and vitexin are flavone 8-C-glycosides that exhibit many biological characteristics. This study aimed to establish a two-enzyme-coupled catalytic strategy to enhance the biosynthesis of orientin and vitexin from apigenin and luteolin, respectively. The C-glucosyltransferase (TcCGT1) gene from Trollius chinensis was cloned and expressed in Escherichia coli BL21(DE3). The optimal activity of TcCGT1 was achieved at pH 9.0 and 37 °C. TcCGT1 was relatively stable over the pH range of 7.0-10.0 at a temperature lower than 45 °C. The coupled catalytic strategy of TcCGT1 and different sucrose synthases was adopted to enhance the production of orientin and vitexin. By optimizing the coupling reaction conditions, orientin and vitexin production successfully achieved 2324.4 and 5524.1 mg/L with a yield of 91.4 and 89.3% (mol/mol), respectively. The coupled catalytic strategy proposed in this study might serve as a promising candidate for the large-scale production of orientin and vitexin in the future.A one-pot rhodium-catalyzed C-H functionalization/organocatalyzed oxa-Michael addition cascade reaction has been developed. This methodology enables the stereodivergent synthesis of diverse 2,3-disubstituted dihydrobenzofurans with broad functional group compatibility in good yields with high levels of stereoselectivity under exceptionally mild conditions. The full complement of stereoisomers of chiral 2,3-disubstituted dihydrobenzofurans and 3,4-disubstituted isochromans could be accessed at will by appropriate permutations of the two chiral catalysts. The current work provides a rare example of two chiral catalysts independently controlling two contiguous stereogenic centers subsequently via a two-step reaction in a single operation.Cosolvent molecular dynamics (CMD) simulations involve an MD simulation of a protein in the presence of explicit water molecules mixed with cosolvent molecules to perform hotspot detection, binding site identification, and binding energy estimation, while other existing methods (e.g., MixMD, SILCS, and MDmix) utilize small molecules that represent functional groups of compounds. However, the cosolvent selections employed in these methods differ and there are only a few cosolvents that are commonly used in these methods. In this study, we proposed a systematic method for constructing a set of cosolvents for drug discovery, termed the EXtended PRObes set construction by REpresentative Retrieval (EXPRORER). First, we extracted typical substructures from FDA-approved drugs, generated 138 cosolvent structures, and for each cosolvent molecule, we conducted CMD simulations to generate a spatial probability distribution map of cosolvent atoms (PMAP). Analyses of PMAP similarity revealed that a cosolvent pair with a PMAP similarity greater than 0.70-0.75 shared similar structural features. We present a method for the construction of a cosolvent subset that satisfies a similarity threshold for all cosolvents, and we tested the constructed sets for four proteins. To our knowledge, this is the first study to include a systematic proposal for cosolvent set construction, and thus, the EXPRORER cosolvents will provide deeper insights into ligand binding sites of various proteins.Extracellular vesicles (EVs) are cell-derived membrane structures that circulate in body fluids and show considerable potential for noninvasive diagnosis. EVs possess surface chemistries and encapsulated molecular cargo that reflect the physiological state of cells from which they originate, including the presence of disease. In order to fully harness the diagnostic potential of EVs, there is a critical need for technologies that can profile large EV populations without sacrificing single EV level detail by averaging over multiple EVs. Here we use a nanofluidic device with tunable confinement to trap EVs in a free-energy landscape that modulates vesicle dynamics in a manner dependent on EV size and charge. As proof-of-principle, we perform size and charge profiling of a population of EVs extracted from human glioblastoma astrocytoma (U373) and normal human astrocytoma (NHA) cell lines.Parkinson's disease (PD) is the second most common neurodegenerative disorder that results in motor dysfunction and, eventually, cognitive impairment. α-Synuclein protein is known as a central protein to the pathophysiology of PD, but the underlying pathological mechanism still remains to be elucidated. In an effort to understand how α-synuclein underlies the pathology of PD, various PD mouse models with α-synuclein overexpression have been developed. However, systemic analysis of the brain proteome of those mouse models is lacking. buy AZD9291 In this study, we established two mouse models of PD by injecting α-synuclein preformed fibrils (PFF) or by inducing overexpression of human A53T α-synuclein to investigate common pathways in the two different types of the PD mouse models. For more accurate quantification of mouse brain proteome, the proteins were quantified using the method of stable isotope labeling with amino acids in mammals . We identified a total of 8355 proteins from the two mouse models; ∼6800 and ∼7200 proteins from α-synuclein PFF-injected mice and human A53T α-synuclein transgenic mice, respectively. Through pathway analysis of the differentially expressed proteins common to both PD mouse models, it was discovered that the complement and coagulation cascade pathways were enriched in the PD mice compared to control animals. Notably, a validation study demonstrated that complement component 3 (C3)-positive astrocytes were increased in the ventral midbrain of the intrastriatal α-synuclein PFF-injected mice and C3 secreted from astrocytes could induce the degeneration of dopaminergic neurons. This is the first study that highlights the significance of the complement and coagulation pathways in the pathogenesis of PD through proteome analyses with two sophisticated mouse models of PD.Despite having the capability to construct benzo-fused heterocycles in complete atom economy and high chemo-, regio-, enantio-, and diastereoselectivities, intramolecular Friedel-Crafts epoxide arene cyclization (IFCEAC) remains underutilized in organic synthesis. The wide adaptation of this powerful Csp2-Csp3 bond-forming reaction, therefore, requires a broad understanding of the substrate scope to better impact heterocycle synthesis. Along this line, we investigated the applicability of IFCEAC for the synthesis of 1,7- and 1,2-fused indoles. In this article, we report the results of our systematic investigation into the scope and limitations of the first examples of the hexafluoro-2-propanol (HFIP)-mediated IFCEAC of readily accessible indolyl-N-tethered epoxides. We observed that the nature and position of the indole and epoxide substituents and the tether length separating these two reacting moieties have strong effects on the cyclization. This mild and transition-metal-free protocol delivered pyrrolo[3,2,1-ij]quinolin-5-ols in moderate to good yields from substrates bearing both a methylene linker that connects the indole and epoxide moieties and an electron-rich indole carbocyclic ring. Notably, the reactions required the presence of a π-activating aryl substituent on the reacting epoxide carbon atom. Interestingly, replacing the methylene tether with an ethylene unit resulted in regioswitching, which delivered the corresponding tetrahydropyrido[1,2-a]indol-8-ols in good to high yields. We could also successfully extend this methodology to pyrrolyl-N-tethered epoxides for a very high-yielding synthesis of tetrahydroindolizin-7-ols.Detonation nanodiamond (DND) is known to form aggregates that significantly reduce their unique nanoscale properties and require postprocessing to separate. How and when DND aggregates is an important question that has not been answered experimentally and could provide the foundation for approaches to limit aggregation. To answer this question, time-resolved small-angle X-ray scattering was performed during the detonation of high-explosives that are expected to condense particulates in the diamond, graphite, and liquid regions of the carbon phase diagram. DND aggregation into low fractal dimension structures could be observed as early as 0.1 μs, along with a separate scattering population also observed from an explosive that produces primarily graphitic products. A counterexample is the case of a high-explosive that produces nano-onions, where no hierarchical scattering was observed for at least 10 μs behind the detonation front. These results suggest that DND aggregation occurs on time scales comparable to particle formation.An unprecedented and expeditious tandem bisannulation of polyfluoroalkylated tetralones with benzamidines to access various fluoroalkyl tetracyclic [1,3]-diazepines through multiple C-N bond formation and C(sp3)-F bond cleavage is reported. The process features high regio-/chemoselectivities, broad substrate scope, good functional group tolerance, procedural simplicity, mild reaction conditions, and scale-up synthesis. Mechanistic studies showed that the distinctive fluorine effect of polyfluoroalkyl tetralone plays a vital role for the aza-tetracycle construction.Dynamic correlation plays an important role in the accurate calculation of chemical compounds such as the description of equilibrium structures in chemical systems. A model for the fast estimation of dynamic correlation energy is introduced in this work. This model is based on the idea of decomposition of the contribution of dynamic correlation energy calculated by nth order Møller-Plesset perturbation (MPn) theory with respect to atomic regions. Multiple levels of theory, including MP2, MP2.5, and MP4, are used as the reference, and the corresponding correlation energy densities are calculated. The proposed model is concise, fast, and promising for practical use, such as the prediction of reaction energies. It can also work as a baseline model or pretrained model for follow-up studies of machine learning.Specialized cellular networks of oxidoreductases coordinate the dithiol/disulfide-exchange reactions that control metabolism, protein regulation, and redox homeostasis. For probes to be selective for redox enzymes and effector proteins (nM to μM concentrations), they must also be able to resist non-specific triggering by the ca. 50 mM background of non-catalytic cellular monothiols. However, no such selective reduction-sensing systems have yet been established. Here, we used rational structural design to independently vary thermodynamic and kinetic aspects of disulfide stability, creating a series of unusual disulfide reduction trigger units designed for stability to monothiols. We integrated the motifs into modular series of fluorogenic probes that release and activate an arbitrary chemical cargo upon reduction, and compared their performance to that of the literature-known disulfides. The probes were comprehensively screened for biological stability and selectivity against a range of redox effector proteins and enzymes. This design process delivered the first disulfide probes with excellent stability to monothiols yet high selectivity for the key redox-active protein effector, thioredoxin. We anticipate that further applications of these novel disulfide triggers will deliver unique probes targeting cellular thioredoxins. We also anticipate that further tuning following this design paradigm will enable redox probes for other important dithiol-manifold redox proteins, that will be useful in revealing the hitherto hidden dynamics of endogenous cellular redox systems.