Removal of signal peptide variants by cation exchange chromatography: A case study.

Signal peptide (SP) is required for secretion of recombinant proteins and typically cleaved by signal peptidase at its C-region to generate the mature proteins. Miscleavage of the SP is reported occasionally, resulting in a truncated- or elongated-terminal sequence. In the present work, we demonstrated that cation exchange (CEX) chromatography is an effective means for removing SP variants with a case study. With the selected resin/conditions, the chromatographic performance is comparable between runs performed at the low end and high end of load density and elution range. The procedure described in this work can be used as a general approach for resin selection and optimization of chromatographic conditions to remove byproducts that bind more strongly than the product to the selected resin.

Comprehensive characterization of the OCT1 phenylalanine-244-alanine substitution reveals highly substrate-dependent effects on transporter function.

Organic cation transporters (OCTs) can transport structurally highly diverse substrates. The molecular basis of this extensive polyspecificity has been further elucidated by cryogenic electron microscopy. Apparently, in addition to negatively charged amino acids, aromatic residues may contribute to substrate binding and substrate selectivity. In this study, we provide a comprehensive characterization of phenylalanine 244 in OCT1 function. We analyzed the uptake of 144 OCT1 substrates for the phenylalanine 244 to alanine substitution compared to wild-type OCT1. This substitution had highly substrate-specific effects ranging from transport reduced to 10% of wild-type activity up to 8-fold increased transport rates. Four percent of substrates showed strongly increased uptake (> 200% of wild type) whereas 39% showed strongly reduced transport (< 50% of wild type). Particularly with larger, more hydrophobic, and more aromatic substrates, the Phe244Ala substitution resulted in higher transport rates and lower inhibition of the transporter. In contrast, substrates with a lower molecular weight and less aromatic rings showed generally decreased uptake rates. A comparison of our data to available transport kinetic data demonstrates that generally, high-affinity low-capacity substrates show increased uptake by the Phe244Ala substitution whereas low-affinity high-capacity substrates are characterized by reduced transport rates. Altogether, our study provides the first comprehensive characterization of the functional role of an aromatic amino acid within the substrate translocation pathway of OCT1. The pleiotropic function further highlights that Phenylalanine 244 interacts in a highly specific manner with OCT1 substrates and inhibitors.

Abiotic Stress Effect on Agastache mexicana subsp. mexicana Yield: Cultivated in Two Contrasting Environments with Organic Nutrition and Artificial Shading.

Research on medicinal plants is essential for their conservation, propagation, resistance to environmental stress, and domestication. The use of organic nutrition has been demonstrated to improve soil fertility and plant quality. It is also important to study the effects of the Basic Cation Saturation Ratio (BCSR) approach, which is a topic where there is currently controversy and limited scientific information. Evaluating the growth and yields of Agastache mexicana subsp. mexicana (Amm) in different environments is crucial for developing effective propagation and domestication strategies. This includes examining warm and subhumid environments with rain in summer in comparison to mild environments with summer rain. Significant differences were observed in the effects of cold, waterlogging, and heat stresses on the plant's biomass yield and the morphometric-quantitative modeling by means of isolines. The biomass yield was 56% higher in environment one compared to environment two, 19% higher in environment one with organic nutrition, and 48% higher in environment two with organic nutrition compared to using only BCSR nutrition. In the second harvesting cycle, the plants in environment one did not survive, while the plants in environment two managed to survive without needing additional nutrition. Statistical and mathematical analyses provided information about the population or sample. Additionally, further analysis using isolines as a new approach revealed new insights into understanding phenology and growth issues.

Hydraulic recharge and element dynamics during salinization in an overexploited coastal aquifer of the world's driest zone: Atacama Desert.

The management of water resources in hyper-arid coastal regions is a challenging task because proper information regarding groundwater recharge and water budget is needed for maintaining the hydraulic balance in optimal conditions, avoiding salinization and seawater intrusion. Thus, this article deals with the estimation of the hydraulic recharge and the study of the effects of salinization on the dynamics of major and trace elements in an alluvial aquifer located in the world's driest zone, the northern Atacama Desert. The result of stable water isotopes (δD and δ18O) and tritium (3H) indicated that groundwater in the area is not recent, whereas 14C results estimated a groundwater residence time ranging between 11,628 and 16,067 yBP. The estimation of the artificial recharge coming from the urban water-supply-system leaks and wastewater/river-water/groundwater infiltration during irrigation was about 19.84 hm3/year, which represents an annual negative water balance of 177 hm3/year for the aquifer. The groundwater salinization triggered by seawater intrusion (up to 32.6 %) has caused the enrichment of Li, Rb, Ca, Ba, and Sr in groundwater by cationic exchange, where the excess of aqueous Na is exchanged by these elements in the aquifer sediments. Other elements such as B, Se, Si, and Sb are enriched in groundwater by ionic strength and/or anionic exchange during salinization. The heightened B concentrations derived from the B-rich alluvial sediments were higher than the limit suggested by international guidelines, representing a risk to consumers. Vanadium seems to be unaffected by salinization, whereas Pb, Mo, As, U, and Zr did not show a clear behavior during saline intrusion. Finally, this article highlights the consequences of conducting improper water management in coastal hyper-arid regions with exacerbated agriculture.

Squid-Inspired Anti-Salt Skin-Like Elastomers With Superhigh Damage Resistance for Aquatic Soft Robots.

Cephalopod skins evolve multiple functions in response to environmental adaptation, encompassing nonlinear mechanoreponse, damage tolerance property, and resistance to seawater. Despite tremendous progress in skin-mimicking materials, the integration of these desirable properties into a single material system remains an ongoing challenge. Here, drawing inspiration from the structure of reflectin proteins in cephalopod skins, a long-term anti-salt elastomer with skin-like nonlinear mechanical properties and extraordinary damage resistance properties is presented. Cation-π interaction is incorporated to induce the geometrically confined nanophases of hydrogen bond domains, resulting in elastomers with exceptional true tensile strength (456.5 ± 68.9 MPa) and unprecedently high fracture energy (103.7 ± 45.7 kJ m-2). Furthermore, the cation-π interaction effectively protects the hydrogen bond domains from corrosion by high-concentration saline solution. The utilization of the resultant skin-like elastomer has been demonstrated by aquatic soft robotics capable of grasping sharp objects. The combined advantages render the present elastomer highly promising for salt enviroment applications, particularly in addressing the challenges posed by sweat, in vivo, and harsh oceanic environments.

Complex Formation of Ag+ and Li+ with Host Molecules Modeled on Intercalation of Graphite.

Pi-stacked and box-shaped host molecules with xanthene as the basis and pyrene as the π-plane were synthesized to verify cation-π interactions between graphene and metal cations. Since crystal structure analysis was not available, DFT calculations were performed to determine the optimized structure, and the π-planes were found to have a slipped parallel structure, with average distances of 456.2-581.0 pm for the stacked compound and 463.4-471.4 pm for the box-shaped compound. Li+ and Ag+ were chosen as acceptors for complexation with metal ions, and their interactions with the π-plane were clarified by NMR titration. Clearly, the interaction with metal ions increased when pyrene π-planes were stacked rather than the pyrene itself. In the stacked compound, the association constants of Ag+ and Li+ were similar; however, in the box-shaped host molecule, only Ag+ had moderate coordination ability, but the interaction with Li+ was very weak, comparable to the interaction with pyrene. As a result, intercalation is more likely to occur in stacked host compound 1, which has some degree of freedom in the pyrene rings, than in the box-shaped compound.

PARAFAC under non-negativity constraint is adapted to recover the underlying Beer-Lambert law of the excitation-emission fluorescence matrix measurements acquired from analyte-triggered semiconductor QDs photoluminescence modulation. When and why?

BACKGROUND: Analyte-triggered semiconductor quantum dots (QDs) modulation in the presence of non-consistently responsive fluorescent species represents a challenging analytical issue in concrete multi-way data handling. QDs with heterogeneous sizes and/or uneven distribution of functional moieties on their surfaces exhibit significant fluctuations in the fluorescent response components, known as chemical rank, across different excitation/emission modes. This phenomenon may lead to a substantial deviation from the proportionality prescribed by Beer-Lambert law. Nonetheless, even in the presence of such deviation, a multi-way model may be successfully selected after determining a proper chemical rank in a QDs system. RESULTS: We show that in a valid PARAllel FACtor (PARAFAC) model under properly determined chemical rank, meaningfully resolved pure spectral profiles can be reached for each fluorescent responsive constituent in the original excitation-emission fluorescence matrix (EEFM) measurements. This was thoroughly illustrated by applying PARAFAC trilinear decomposition of a three-way data array of two distinct datasets acquired from semiconductor QDs sensing systems with low-rank trilinear assumption. The first dataset, presented here for the first time, comprises EEFM measurements of the ligand-driven quenching of thiomalic acid (TMA)-capped AgInS2 (AIS) QDs by vomitoxin. The second dataset, employed for illustrative purposes, comprises EEFM measurements of the quenching, via cation bridging, of glutathione (GSH)-capped CdTe QDs by Pb(II). The results of this study enabled the determination of vomitoxin at a ppb level in real samples of fish feeds, showcasing the efficacy of the PARAFAC model in resolving spectral signatures (loadings) and pure concentration profiles (scores). SIGNIFICANCE: PARAFAC under a properly examined chemical rank can be easily adapted for retrieval the underlying Beer-Lambert law of the original EEFM measurements with a low-rank trilinear structure through the chemically meaningful information either when (i) no deviation of Beer-Lambert law was observed as deeply discussed in connection with the dataset acquired from vomitoxin-driven molecular sensing through TMA-capped AIS QDs, or when (ii) substantial deviations of the Beer-Lambert law are evident, as discussed in connection with the dataset collected from sensing ionic species through Pb(II) bridging of GSH-capped CdTe QDs.

The high-light-sensitivity mechanism and optogenetic properties of the bacteriorhodopsin-like channelrhodopsin GtCCR4.

Channelrhodopsins are microbial light-gated ion channels that can control the firing of neurons in response to light. Among several cation channelrhodopsins identified in Guillardia theta (GtCCRs), GtCCR4 has higher light sensitivity than typical channelrhodopsins. Furthermore, GtCCR4 shows superior properties as an optogenetic tool, such as minimal desensitization. Our structural analyses of GtCCR2 and GtCCR4 revealed that GtCCR4 has an outwardly bent transmembrane helix, resembling the conformation of activated G-protein-coupled receptors. Spectroscopic and electrophysiological comparisons suggested that this helix bend in GtCCR4 omits channel recovery time and contributes to high light sensitivity. An electrophysiological comparison of GtCCR4 and the well-characterized optogenetic tool ChRmine demonstrated that GtCCR4 has superior current continuity and action-potential spike generation with less invasiveness in neurons. We also identified highly active mutants of GtCCR4. These results shed light on the diverse structures and dynamics of microbial rhodopsins and demonstrate the strong optogenetic potential of GtCCR4.

Cation Vacancy Strategy Activated Layered Double Hydroxides: Enhanced Electrochemical Performance and Longevity.

The defect design strategy has been extensively employed to enhance the reaction kinetics of layered double hydroxide (LDH) electrode materials. Furthermore, it is anticipated to improve the cyclic stability of LDHs through this approach, serving a dual purpose. However, the potential mechanisms of cation vacancies' impact on the electrochemical performance of electrodes at the atomic scale still need further clarification. In this study, a typical aluminum-vacancy LDH material via a simple alkaline etching method was demonstrated. Electrochemical in situ Raman spectroscopy, ex situ X-ray diffraction (XRD), and first-principles calculations were utilized to elucidate the mechanism of Faradaic reactions. The findings indicate that this cation vacancy strategy not only enhances the electrochemical reaction kinetics of the electrodes but also effectively reduces the energy barrier for the α to γ phase transition of LDHs during the charge-discharge processes, thereby enhancing its longevity. To further validate the practical application of this defect design, an asymmetric solid-state supercapacitor was formed, which maintains 93.9% capacity after 20 000 charge-discharge cycles. This research offers technical guidance for the development of a new generation of high-performance and long-life LDH electrode materials based on a cation vacancy strategy.

Alkali metal cations enhance CO2 reduction by a Co molecular complex in a bipolar membrane electrolyzer.

The electrochemical reduction of CO2 is a promising pathway for converting CO2 into valuable fuels and chemicals. The local environment at the cathode of CO2 electrolyzers plays a key role in determining activity and selectivity, but currently some mechanisms are still under debate. In particular, alkali metal cations have been shown to enhance the selectivity of metal catalysts, but their role remains less explored for molecular catalysts especially in high-current electrolyzers. Here, we investigated the enhancement effects of cations (Na+, K+, Cs+) on Co phthalocyanine (CoPc) in a state-of-the-art reverse-biased bipolar membrane electrolyzer. When added to the anolyte, these cations increased the Faradaic efficiency for CO, except in the case of Na+ in which the effect was transient, but the effects are convoluted with the transport process through the membrane. Alternatively, these cations can also be added directly to the cathode as chloride salts, allowing the use of a pure H2O anolyte feed, leading to sustained improved CO selectivity (61% at 100 mA cm-2 after 24 h). Our results show that cation addition is a simple yet effective strategy for improving the product selectivity of molecular electrocatalysts, opening up new avenues for tuning their local environment for CO2 reduction.This article is part of the discussion meeting issue 'Green carbon for the chemical industry of the future'.

Covalent Organic Framework Membranes with Regulated Orientation for Monovalent Cation Sieving.

Continuous covalent organic framework (COF) thin membranes have garnered broad concern over the past few years due to their merits of low energy requirements, operational simplicity, ecofriendliness, and high separation efficiency in the application process. This study marks the first instance of fabricating two distinct, self-supporting COF membranes from identical building blocks through solvent modulation. Notably, the precision of the COF membrane's separation capabilities is substantially enhanced by altering the pore alignment from a random to a vertical orientation. Within these confined channels, the membrane with vertically aligned pores and micron-scale stacking thickness demonstrates rapid and selective transportation of Li+ ions over Na+ and K+ ions, achieving Li+/K+ and Li+/Na+ selectivity ratios of 38.7 and 7.2, respectively. This research not only reveals regulated orientation and layer stacking in COF membranes via strategic solvent selection but also offers a potent approach for developing membranes specialized in Li+ ion separation.

Organic cation transporter 1 in the lacrimal gland facilitates the entry of systemic drugs causing ocular surface toxicity.

Many of the daily systemic medications (parenteral and oral) used to treat various diseases are known to cause ocular toxicities - leading to vision loss. How these medications gain entry into the eye despite the ocular barriers is an important question to be addressed. Various reports show almost 30-40 % of systemic drugs causing ocular toxicity are organic cation in nature. We hypothesize these systemic drugs (cations) are non-specifically recognized as endogenous substrates by organic cation transporter (OCT1) in the lacrimal gland, thereby facilitating its entry into the anterior eye segment. Therefore, we studied the expression and localization of OCT1 in the lacrimal gland of rabbits. Further, to prove our hypothesis, OCT1 substrates (known as well as predicted from our previous Artificial Intelligence study) were administered intravenously in the presence and absence of topically administered OCT1 blockers. Our findings show, OCT1 gene and protein expression in the lacrimal gland, with its localization in the terminal acinar cells. The tear levels of intravenously administered substrates decreased in the presence of topical OCT1 blockers, indicating - a) the entry of systemic drugs into the eye via lacrimal secretion and b) OCT1 in the lacrimal gland is involved in the drug transport (substrates) from blood to the eye. Though the role of transporters in toxicity is well-known, the current study opens a new avenue for understanding the role of transporters in ocular toxicity.

Analysis of Ca1-xSrxCO3 phases generated by competitive Sr2+ replacement in pre-formed aragonite.

The ratio of Sr/Ca ions in marine biogenic minerals is considered advantageous for tracking geochemical and biomineralization processes that occur in the oceans. It is debatable, though, whether the ratio in biominerals such as coral skeleton is simply related to values in the seawater environment or controlled by the organism. Recent data show that coral larvae produce partially disordered immature aragonite in Mg-containing Sr-poor calcifying fluids, which transforms into well-ordered aragonite in Mg-depleted Sr-enriched environments, upon animal metamorphosis into the sessile polyp state. Inspired by the process in young coral, we explored in vitro substitution of Ca by Sr in aragonite by exposing aragonite crystals precipitated a priori to Sr solutions with variable concentrations. The resulting biphasic material, comprised of Sr-doped aragonite and Ca-doped strontianite, was carefully analyzed for foreign cation substitution in each polymorph. This allowed to establish a linear correlation between Sr levels in mineralizing solutions and Sr in aragonite as well as Ca in strontianite. It indicated that ca. 5-fold higher Sr solution concentration is needed for substitution in the crystal to reach the level found in corals. It also provided with Sr levels required for a putative strontianite phase to form.

Effect of Cations on ATP Binding to the N-domain of Na+, K+-ATPase.

The nucleotide-binding domain (N-domain) of the Na+, K+-ATPase (NKA) is physicochemically characterized by a high content of Glu and Asp residues, resulting in a low isoelectric point (pI = 5.0). Acidic proteins are known to interact with cations. The analysis in silico revealed potential cation interaction sites in the NKA N-domain structure. The interaction with cations was tested in vitro by using a recombinant NKA N-domain. The N-domain contains two Trp residues at the protein surface, as determined by acrylamide-mediated fluorescence quenching, that are useful for structural studies through fluorescence changes. Intrinsic fluorescence of the N-domain was decreased by the presence of cations (Na+, K+, Ca2+) indicating an effect on the protein structure. ATP binding also decreased the N-domain intrinsic fluorescence, which allowed nucleotide affinity determination. In the presence of cations, the N-domain affinity for ATP was increased. Molecular docking of fluorescein isothiocyanate (FITC) with the N-domain showed two binding modes with the isothiocyanate group located 5-6 Å close to Lys485 and Lys506 in the nucleotide-binding site. The presence of ATP prevented the FITC covalent labeling of the N-domain demonstrating the competitive behavior for the binding site. It is proposed that cations interact with the N-domain structure and thereby modulate nucleotide (ATP) affinity and possibly affecting NKA catalysis.

Construction of a triple-mode sensing platform for effective differentiation, speciation, and monitoring of mercury and methylmercury bioaccumulation in crayfish via cation exchange reaction.

Developing rapid and sensitive methods for monitoring inorganic mercury (Hg2+) and methylmercury (CH3Hg+) in crayfish is crucial for understanding the environmental impact of these contaminants. In this work, a novel tri-mode strategy was developed for highly sensitive monitoring of Hg2+ and CH3Hg+ bioaccumulation in crayfish by inductively coupled plasma mass spectrometry (ICP-MS)/ fluorescence /smartphone colorimetric (RGB) analysis without chromatographic separation. Cation exchange reaction (CER) was performed between Hg2+ and luminescent CdTe quantum dots (QDs), while CH3Hg+ unrealizable CER. The CH3Hg+ can be transformed to Hg2+ by simple UV irradiation, speciation analysis can be realized by detecting the fluorescence of CdTe QDs after incubation by Hg2+ and total Hg2+. In addition, the filtration of reacted CdTe QDs was carried out, ICP-MS was performed to detect exchanged Cd2+ by Hg2+ and total Hg2+, as well the smartphone RGB analysis was performed for membrane colorimetry. The limits of detection (LODs) of Hg2+ and CH3Hg+ for ICP-MS, fluorescence, and colorimetric (RGB) modes were 0.03 ng mL-1, 18 ng mL-1, and 0.9 µg mL-1 respectively. Density Functional Theory (DFT) was employed to validate the mechanism of the CER reaction. CdTe QDs array analysis with five different ligands was performed to eliminate potential ion interferences of Ag+ and Cu2+ that could occur during the CER reaction. The well-designed system was successfully utilized for monitoring trace Hg2+ and CH3Hg+ in crayfish fed Hg2+ and CH3Hg+ contaminative food over a two-week "uptake" period and a three-week "depuration" period. The results indicated that the Hg2+ uptake in different tissues was significantly different from that of CH3Hg+ in all tissues. There was evidence of Hg uptake from water via leaching from food, although the principal source of uptake was from food.

Interaction of heavy metals and polycyclic aromatic hydrocarbons in soil-crop systems: The effects and mechanisms.

In natural environments, the removal and degradation of two major pollutants, heavy metals (HMs) and polycyclic aromatic hydrocarbons (PAHs), are explored through targeted experimental investigations. However, these endeavors reveal that outcomes in situ may significantly diverge from the idealized effects observed in laboratory settings due to the complex interaction between HMs and PAHs, underscoring a pressing need for thorough research into their mutual impacts. This review examines the origins and migratory pathways of compound pollution stemming from HMs and PAHs. Concurrently, it provides an overview of the farmland ecosystem's response to combined HMs-PAHs pollution. This encompassed the assessments of changes in the soil's physical and chemical properties, the intricacies of the migration and transformation processes of the combined pollution within plants, and the consequential impact on the physiological functions of soil microorganisms. The varying concentration ratios of HMs and PAHs can modulate the permeability of plant root cell membranes, thereby influencing the translocation of these substances within the plant via symplastic and apoplastic pathways. Recent research has uncovered the mechanisms underlying cation-π interactions between HMs and PAHs. This review aims to offer a comprehensive overview of the current state of HMs-PAHs co-pollution, offering both qualitative and quantitative insights into their interaction patterns within the farmland ecosystem. The ultimate goal is to establish a robust theoretical foundation to support the in-situ remediation of these pollutants in agricultural practices and to provide a theoretical basis for soil health management in agricultural production.

Unveiling the Cation Dependence in Alkaline Hydrogen Evolution by Differently-Charged Ruthenium/Molybdenum Sulfide Hybrids.

The sluggish kinetics of hydrogen evolution reaction (HER) via water reduction limits the efficiency of alkaline water electrolysis. The HER kinetics is not only intimately related to the catalyst surface structure but also relevant to the cation identity of the electrolyte. The cation dependence also relies on the surface electronic structure and applied potential, but this interrelated effect and its underlying mechanism awaits elucidation. Herein, differently-charged molybdenum sulfide (MoSx) cluster supports ([Mo3S13]2- and [Mo3S7]4+) are utilized to hybridize with the identical metallic Ru centers. The specific electrostatic interaction between MoSx clusters and Ru precursors induces different Ru valences of the hybrids, with a higher valence state for Ru/Mo3S13 endowing a higher activity. The Ru/Mo3S13 and Ru/Mo3S7 exhibited drastically-different cation dependence, in which the charged support determines the local accumulation of cations and resulting water structures. The more negatively-charged Mo3S13 support induces the facile accumulation of cations, especially for less-hydrated K+ cations. The water activation capability by Ru valences and cation accumulation from the support effect in-together determine the cation-dependent alkaline HER activity. This work not only enriches the understanding about the cation-dependent HER mechanism but also shines a light on the rational optimization strategy of electrode/electrolyte interfaces.

One-Pot Transition-Metal-Free Synthesis of Alkynyl Amides.

Alkynyl amides play crucial roles in organic synthesis in the production of bioactive compounds and valuable heterocycles. Despite numerous studies on their synthesis, challenges persist due to the necessity of harsh or hazardous conditions and the use of costly or unstable reagents. Herein, we present a one-pot method for the synthesis of all three bonds of the alkyne under transition-metal free conditions. An important feature of this chemistry is the use of readily available feedstock chemicals, such as methyl esters and acetamides. This approach offers efficient access to a wide range of aryl and alkyl alkynyl amides and demonstrates excellent tolerance towards various functional groups in a sustainable and cost-effective manner.

HcZnT2 is a highly mycorrhiza-induced zinc transporter from Hebeloma cylindrosporum in association with pine.

Zinc (Zn) shortage is a common micronutrient deficiency affecting plants worldwide, while Zn toxicity may occur when this metal is in excess. Ectomycorrhizal (ECM) fungi are known to be able to modulate the transfer of macro- and microelements, among them Zn, to the plant. However, the underlying mechanisms are not well understood. We identified the HcZnT2 gene from the ECM fungus Hebeloma cylindrosporum, encoding a member of the Cation Diffusion Facilitator (CDF) family including Zn transporters, and analyzed its transcriptional regulation, the transport function by yeast complementation experiments, and its subcellular localization using a GFP fusion protein in yeast. HcZnT2 is highly induced during mycorrhization of Pinus pinaster, and upregulated in presence of the host plant root even without any direct contact. However, HcZnT2 is repressed by Zn excess conditions. By functional expression in yeast, our results strongly support the ability of HcZnT2 to transport Zn and, to a lesser extent, manganese. HcZnT2 localization was associated with the endoplasmic reticulum of yeast. Mycorrhizal gene activation at low external Zn suggests that the Zn transporter HcZnT2 might be important for the early establishment of the ECM symbiosis during Zn deficiency, rather than under Zn excess. HcZnT2 arises as an extremely remarkable candidate playing a key role in Zn homeostasis and regulation in ectomycorrhiza.

Inhibitory effects of flavonoids on organic cation transporter 1: Implications for food/herb-drug interactions and hepatoprotective effects.

Organic cation transporter 1 (OCT1, gene symbol: SLC22A1) is mainly responsible for the hepatic uptake of various cationic drugs, closely associated with drug-induced liver injury (DILI). Screening and identifying potent OCT1 inhibitors with little toxicity in natural products is of great value in alleviating OCT1-mediated liver injury. Flavonoids, a group of polyphenols commonly found in foodstuffs and herbal products, have been reported to cause transporter-mediated food/herb-drug interactions (FDIs). Our objective was to investigate potential inhibitors of OCT1 from 96 flavonoids, evaluate the hepatoprotective effects on retrorsine-induced liver injury, and clarify the structure-activity relationships of flavonoids with OCT1. Thirteen flavonoids exhibited significant inhibition (>50%) on OCT1 in OCT1-HEK293 cells. Among them, the five strongest flavonoid inhibitors (IC50 < 10 µM), including α-naphthoflavone, apigenin, 6-hydroxyflavone, luteolin, and isosilybin markedly decreased oxaliplatin-induced cytotoxicity. In retrorsine-induced liver injury models, they also reduced alanine aminotransferase (ALT) and aspartate aminotransferase (AST) to different levels, the best of which was 6-hydroxyflavone. The pharmacophore model clarified that hydrogen bond acceptors at the 4,8,5' position might play a vital role in the inhibitory effect of flavonoids on OCT1. Taken together, our findings would pave the way to predicting the potential risks of flavonoid-related FDIs in humans and optimizing flavonoid structure to alleviate OCT1-mediated liver injury.