Oxalate modification enabled advanced phosphate removal of nZVI: In Situ formed surface ternary complex and altered multi-stage adsorption process.

Nano zero-valent iron (nZVI) is a promising phosphate adsorbent for advanced phosphate removal. However, the rapid passivation of nZVI and the low activity of adsorption sites seriously limit its phosphate removal performance, accounting for its inapplicability to meet the emission criteria of 0.1 mg P/L phosphate. In this study, we report that the oxalate modification can inhibit the passivation of nZVI and alter the multi-stage phosphate adsorption mechanism by changing the adsorption sites. As expected, the stronger anti-passivation ability of oxalate modified nZVI (OX-nZVI) strongly favored its phosphate adsorption. Interestingly, the oxalate modification endowed the surface Fe(III) sites with the lowest chemisorption energy and the fastest phosphate adsorption ability than the other adsorption sites, by in situ forming a Fe(III)-phosphate-oxalate ternary complex, therefore enabling an advanced phosphate removal process. At an initial phosphate concentration of 1.00 mg P/L, pH of 6.0 and a dosage of 0.3 g/L of adsorbents, OX-nZVI exhibited faster phosphate removal rate (0.11 g/mg/min) and lower residual phosphate level (0.02 mg P/L) than nZVI (0.055 g/mg/min and 0.19 mg P/L). This study sheds light on the importance of site manipulation in the development of high-performance adsorbents, and offers a facile surface modification strategy to prepare superior iron-based materials for advanced phosphate removal.

When Metal Complexes Evolve, and a Minor Species Is the Most Active: the Case of Bis(Phenanthroline)Copper in the Catalysis of Glutathione Oxidation and Hydroxyl Radical Generation.

Several copper-ligands, including 1,10-phenanthroline (Phen), have been investigated for anticancer purposes based on their capacity to bind excess copper (Cu) in cancer tissues and form redox active complexes able to catalyse the formation of reactive oxygen species (ROS), ultimately leading to oxidative stress and cell death. Glutathione (GSH) is a critical compound as it is highly concentrated intracellularly and can reduce and dissociate copper(II) from the ligand forming poorly redox-active copper(I)-thiolate clusters. Here we report that Cu-Phen2  speciation evolves in physiologically relevant GSH concentrations. Experimental and computational experiments suggest that at pH 7.4 mostly copper(I)-GSH clusters are formed, but a minor species of copper(I) bound to one Phen and forming ternary complexes with GSH (GS-Cu-Phen) is the redox active species, oxidizing quite efficiently GSH to GSSG and forming HO• radicals. This minor active species becomes more populated at lower pH, such as typical lysosomal pH 5, resulting in faster GSH oxidation and HO• production. Consistently, cell culture studies showed lower toxicity of Cu-Phen2 upon inhibition of lysosomal acidification. Overall, this study underscores that sub-cellular localisation can considerably influence the speciation of Cu-based drugs and that minor species can be the most redox- and biologically- active.

Secondary Sphere Lewis Acid Activated Heme Superoxo Adducts Mimic Crucial Non-Covalent Interactions in IDO/TDO Heme Dioxygenases.

Heme enzymes play a central role in a medley of reactivities within a wide variety of crucial biological systems. Their active sites are highly decorated with pivotal evolutionarily optimized non-covalent interactions that precisely choreograph their biological functionalities with specific regio-, stereo-, and chemo-selectivities. Gaining a clear comprehension of how such weak interactions within the active sites control reactivity offers powerful information to be implemented into the design of future therapeutic agents that target these heme enzymes. To shed light on such critical details pertaining to tryptophan dioxygenating heme enzymes, this study investigates the indole dioxygenation reactivities of Lewis acid-activated heme superoxo model systems, wherein an unprecedented kinetic behavior is revealed. In that, the activated heme superoxo adduct is observed to undergo indole dioxygenation with the intermediacy of a non-covalently organized precursor complex, which forms prior to the rate-limiting step of the overall reaction landscape. Spectroscopic and theoretical characterization of this precursor complex draws close parallels to the ternary complex of heme dioxygenases, which has been postulated to be of crucial importance for successful 2,3-dioxygenative cleavage of indole moieties.

LFHP-1c improves cognitive function after TBI in mice by reducing oxidative stress through the PGAM5-NRF2-KEAP1 ternary complex.

Traumatic brain injury (TBI) is a leading cause of disability and death. Thus, timely and effective secondary brain injury intervention is crucial, with potential to improve the prognosis of TBI. Oxidative stress contributes to post-traumatic secondary cognitive impairment, and the reduction of post-traumatic oxidative stress effectively enhances cognitive function. Phosphoglycerate-mutating enzyme 5 (PGAM5), a member of the phosphoglycerate transporter enzyme family, is upregulated in TBI and induces mitochondrial autophagy. This further exacerbates damage following TBI. The present study focused on the small molecule drug, LFHP-1c, which is a novel inhibitor of PGAM5. The present study used an in vivo mouse model incorporating a controlled cortical impact-induced TBI, to examine the impact of LFHP-1c on oxidative stress and cognitive function. The present study aimed to determine the impact of LFHP-1c on the PGAM5-Kelch-like ECH-associated protein 1 (KEAP1)- nuclear factor erythroid 2-related factor 2 (NRF2) ternary complex within the TBI context. Results of the present study indicated that LFHP-1c suppresses PGAM5 expression and inhibits the development of the PGAM5-KEAP1-NRF2 ternary complex, thereby promoting the release of NRF2 and KEAP1. This in turn promotes the entry of NRF2 into the nucleus following TBI, leading to increased expression of anti-oxidative stress downstream factors, such as heme oxygenase-1, glutathione peroxidase 1 and superoxide dismutase 1. In addition, LFHP-1c also released KEAP1, leading to mitochondrial Rho GTPase 2 degradation and reducing perinuclear aggregation of mitochondria in the cell, which reduced oxidative stress and ultimately improved cognitive function after TBI.

Leveraging Covalency to Stabilize Ternary Complex Formation For Cell-Cell "Induced Proximity".

Recent advances in the field of translational chemical biology use diverse "proximity-inducing" synthetic modalities to elicit new modes of "event driven" pharmacology. These include mechanisms of targeted protein degradation and immune clearance of pathogenic cells. Heterobifunctional "chimeric" compounds like Proteolysis TArgeting Chimeras (PROTACs) and Antibody Recruiting Molecules (ARMs) leverage these mechanisms, respectively. Both systems function through the formation of reversible "ternary" or higher-order biomolecular complexes. Critical to function are key parameters, such as bifunctional molecule affinity for endogenous proteins, target residence time, and turnover. To probe the mechanism and enhance function, covalent chemical approaches have been developed to kinetically stabilize ternary complexes. These include electrophilic PROTACs and Covalent Immune Recruiters (CIRs), the latter designed to uniquely enforce cell-cell induced proximity. Inducing cell-cell proximity is associated with key challenges arising from a combination of steric and/or mechanical based destabilizing forces on the ternary complex. These factors can attenuate the formation of ternary complexes driven by high affinity bifunctional/proximity inducing molecules. This Account describes initial efforts in our lab to address these challenges using the CIR strategy in antibody recruitment or receptor engineered T cell model systems of cell-cell induced proximity. ARMs form ternary complexes with serum antibodies and surface protein antigens on tumor cells that subsequently engage immune cells via Fc receptors. Binding and clustering of Fc receptors trigger immune cell killing of the tumor cell. We applied the CIR strategy to convert ARMs to covalent chimeras, which "irreversibly" recruit serum antibodies to tumor cells. These covalent chimeras leverage electrophile preorganization and kinetic effective molarity to achieve fast and selective covalent engagement of the target ternary complex protein, e.g., serum antibody. Importantly, covalent engagement can proceed via diverse binding site amino acids beyond cysteine. Covalent chimeras demonstrated striking functional enhancements compared to noncovalent ARM analogs in functional immune assays. We revealed this enhancement was in fact due to the increased kinetic stability and not concentration, of ternary complexes. This finding was recapitulated using analogous CIR modalities that integrate peptidic or carbohydrate binding ligands with Sulfur(VI) Fluoride Exchange (SuFEx) electrophiles to induce cell-cell proximity. Mechanistic studies in a distinct model system that uses T cells engineered with receptors that recognize covalent chimeras or ARMs, revealed covalent receptor engagement uniquely enforces downstream activation signaling. Finally, this Account discusses potential challenges and future directions for adapting and optimizing covalent chimeric/bifunctional molecules for diverse applications in cell-cell induced proximity.

Mechanism of sucrose improving the mechanical characteristics of foams stabilized by soy protein isolate/gellan gum/guar gum ternary complex.

Sucrose shows the potential of stabilizing foam system. This study systematically evaluated the mechanism by which sucrose improved foaming properties and mechanical characteristics of foams stabilized by soy protein isolate/gellan gum/guar gum ternary complex. Results showed that sucrose could bond to the surface of ternary complex or self-aggregate within the continuous phase, resulting in the neutralization of charges (nearly zero) and an increase in particle size (up to 62.54 µm). The addition of 30 % sucrose reinforced foam system with an increased foamability (305.99 %) but a longer foaming time (10 min) during foaming process. Moreover, the mechanical characteristics, including hardness, elastic strength (Power-law constant) and solid characteristic (frequency exponent), were also significantly enhanced to 1.26 N, 354.7956 and 2.5873, respectively, which were 1.65, 1.94 and 1.11 times than those of foams without sucrose. The microscopic mechanism lied in the reduced water freedom degree caused by sucrose, which generated a compact structural network around bubbles for providing a stable and stiff structure to foams. These findings will provide clear theoretical guidance for regulating mechanical characteristics of aerated foods by using sucrose as structural building blocks.

Regulated induced proximity targeting chimeras-RIPTACs-A heterobifunctional small molecule strategy for cancer selective therapies.

We describe a protein proximity inducing therapeutic modality called Regulated Induced Proximity Targeting Chimeras or RIPTACs: heterobifunctional small molecules that elicit a stable ternary complex between a target protein (TP) selectively expressed in tumor cells and a pan-expressed protein essential for cell survival. The resulting co-operative protein-protein interaction (PPI) abrogates the function of the essential protein, thus leading to death selectively in cells expressing the TP. This approach leverages differentially expressed intracellular proteins as novel cancer targets, with the advantage of not requiring the target to be a disease driver. In this chemical biology study, we design RIPTACs that incorporate a ligand against a model TP connected via a linker to effector ligands such as JQ1 (BRD4) or BI2536 (PLK1) or CDK inhibitors such as TMX3013 or dinaciclib. RIPTACs accumulate selectively in cells expressing the HaloTag-FKBP target, form co-operative intracellular ternary complexes, and induce an anti-proliferative response in target-expressing cells.

Construction of sodium alginate/trehalose/wheat starch ternary complex and its effects on storage stability of frozen dough system.

In order to improve the quality of frozen dough, a calcium alginate-coated sodium alginate/trehalose/wheat starch ternary complex was designed in this paper. The ternary complex was added to dough, and the dough quality were measured after 0-30 d of frozen storage. The XRD and FT-IR results showed the ternary complex was mainly starchy crystal. The TGA curves showed the starting (To), peak (Tp) and termination temperature (Tc) were increased. The interaction between sodium alginate and trehalose enhanced the thermal performance of ternary complex. As the ternary complex addition to dough increased, the maximum ice crystal formation zone of the frozen dough passed faster, resulting in more uniform and smaller ice crystals. The dough with 0.8% addition contained more bound water and had better hardness, springiness and cohesiveness. In conclusion, the study provides a novel insight and understanding for the development of ternary complex as food additives in frozen food industry.

Quantitative Insights into Phosphate-Enhanced Lead Immobilization on Goethite.

Despite extensive study, geochemical modeling often fails to accurately predict lead (Pb) immobilization in environmental samples. This study employs the Charge Distribution MUlti-SIte Complexation (CD-MUSIC) model, X-ray absorption fine structure (XAFS), and density functional theory (DFT) to investigate mechanisms of phosphate (PO4) induced Pb immobilization on metal (hydr)oxides. The results reveal that PO4 mainly enhances bidentate-adsorbed Pb on goethite via electrostatic synergy at low PO4 concentrations. At relatively low pH (below 5.5) and elevated PO4 concentrations, the formation of the monodentate-O-sharing Pb-PO4 ternary structure on goethite becomes important. Precipitation of hydropyromorphite (Pb5(PO4)3OH) occurs at high pH and high concentrations of Pb and PO4, with an optimized log Ksp value of -82.02. The adjustment of log Ksp compared to that in the bulk solution allows for quantification of the overall Pb-PO4 precipitation enhanced by goethite. The CD-MUSIC model parameters for both the bidentate Pb complex and the monodentate-O-sharing Pb-PO4 ternary complex were optimized. The modeling results and parameters are further validated and specified with XAFS analysis and DFT calculations. This study provides quantitative molecular-level insights into the contributions of electrostatic enhancement, ternary complexation, and precipitation to phosphate-induced Pb immobilization on oxides, which will be helpful in resolving controversies regarding Pb distribution in environmental samples.

eIF2ß zinc-binding domain interacts with the eIF2γ subunit through the guanine nucleotide binding interface to promote Met-tRNAiMet binding.

The heterotrimeric eIF2 complex consists of a core eIF2γ subunit to which binds eIF2α and eIF2ß subunits and plays an important role in delivering the Met-tRNAiMet to the 40S ribosome and start codon selection. The intricacies of eIF2ß-γ interaction in promoting Met-tRNAiMet binding are not clearly understood. Previously, the zinc-binding domain (ZBD) eIF2ßS264Y mutation was reported to cause Met-tRNAiMet binding defect due to the intrinsic GTPase activity. We showed that the eIF2ßS264Y mutation has eIF2ß-γ interaction defect. Consistently, the eIF2ßT238A intragenic suppressor mutation restored the eIF2ß-γ and Met-tRNAiMet binding. The eIF2ß-ZBD residues Asn252Asp and Arg253Ala mutation caused Met-tRNAiMet binding defect that was partially rescued by the eIF2ßT238A mutation, suggesting the eIF2ß-ZBD modulates Met-tRNAiMet binding. The suppressor mutation rescued the translation initiation fidelity defect of the eIF2γN135D SW-I mutation and eIF2ßF217A/Q221A double mutation in the HTH domain. The eIF2ßT238A suppressor mutation could not rescue the eIF2ß binding defect of the eIF2γV281K mutation; however, combining the eIF2ßS264Y mutation with the eIF2γV281K mutation was lethal. In addition to the previously known interaction of eIF2ß with the eIF2γ subunit via its α1-helix, the eIF2ß-ZBD also interacts with the eIF2γ subunit via guanine nucleotide-binding interface; thus, the eIF2ß-γ interacts via two distinct binding sites.

Evaluating the binding mechanism, structural changes and stability of ternary complexes formed by the interaction of folic acid with whey protein concentrate-80 and L-ascorbyl 6-palmitate.

In the present study, we investigated the mechanisms associated with the stabilizing effects of whey protein concentrate-80 (WPC80) and L-ascorbyl 6-palmitate (LAP) on folic acid (FA). Multispectral techniques show that WPC80 binds to FA and LAP mainly through hydrophobic interactions, and that energy is transferred from WPC80 to FA and LAP in a nonradiative form (FA/LAP); The combination of FA/LAP resulted in a change in the conformation and secondary structure content of WPC80, an increase in the absolute zeta potential of the system, and a shift in the particle size distribution towards smaller sizes. The compound system exhibits strengthened antioxidant properties and favorable binding properties. Besides, WPC80 improves the storage stability of FA under different conditions. These results demonstrated that the ternary complex formed by FA co-binding with WPC80 and LAP is an effective way to improve the stability against of FA.

Single Effective Complex Loading into Zero-Mode Waveguides Optimized with Fluorescence Evaluation at Quenching and Accumulation Checkpoints.

Single-molecule detection with high accuracy and specialty plays an important role in biomedical diagnosis and screening. Zero-mode waveguides (ZMWs) enable the possibility of single biological molecule detection in real time. Nevertheless, the absence of a reliable assessment for single effective complex loading has constrained further applications of ZMWs in complex interaction. Both the quantity and activity of the complex loaded into ZMWs have a critical effect on the efficiency of detection. Herein, a fluorescence evaluation at quenching and accumulation checkpoints was established to assess and optimize single effective complex loading into ZMWs. A primer-template-enzyme ternary complex was designed, and then an evaluation for quantity statistics at the quenching checkpoint and functional activity at the accumulation checkpoint was used to validate the effectiveness of complexes loaded into ZMWs. By optimizing the parameters such as loading time, procedures, and enzyme amount, the single-molecule effective occupancy was increased to 25.48%, achieving 68.86% of the theoretical maximum value (37%) according to Poisson statistics. It is of great significance to provide effective complex-loading validation for improving the sample-loading efficiency of single-molecule assays or sequencing in the future.

The impact of addition of pearl millet starch-germ complex in white bread on nutritional, textural, structural, and glycaemic response: Single blinded randomized controlled trial in healthy and pre-diabetic participants.

The rise of pre-diabetes at the global level has created a significant interest in developing low glycaemic index food products. The pearl millet is a cheaper source of starch and its germ contains significant amount of protein and fat. The complexing of pearl millet starch and germ by dry heat treatment (PMSGH) resulted an increase in the resistant starch content upto 45.09 % due to formation of amylose-glutelin-linoleic acid complex. The resulting pearl millet starch germ complex was incorporated into wheat bread at 20, 25, and 30 %. The PMSGH incorporated into bread at 30 % reduced the glycaemic index to 52.31. The PMSGH incorporated bread had significantly (p < 0.05)increased in the hardness with a reduction in springiness and cohesiveness. The structural attributes of the 30 % PMSGH incorporated bread revealed a significant (p < 0.05)increase in 1040/1020 cm-1 ratio and relative crystallinity. The consumption of functional bread incorporated with pearl millet starch germ complex reduced blood glucose levels and in vivo glycaemic index in healthy and pre-diabetic participants when compared to white bread. Hence, the study showed that the incorporation of pearl millet starch-germ complex into food products could be a potential new and healthier approach for improving dietary options in pre-diabetes care.

Effect of a novel siRNA delivery system, siRNA ternary complex, on melanoma lung metastasis.

In this study, we determined effects of an anionic siRNA delivery vector, siRNA ternary complex, which is constructed with biodegradable dendrigraft poly-L-lysine (DGL) and γ-polyglutamic acid (γ-PGA) on the melanoma cells and melanoma lung metastasis. The siRNA ternary complex showed high cellular uptake and silencing effect in melanoma cell line B16-F10/Luc cells. After intravenous administration of the siRNA ternary complex, high silencing effect was also observed in the lung of B16-F10/Luc melanoma metastasis model mice. Therefore, we applied vascular endothelial growth factor (VEGF)-siRNA on the siRNA ternary complex and determined the effect on the melanoma lung metastasis. The siRNA ternary complex containing VEGF-siRNA reduced VEGF protein levels significantly in in vitro and in vivo, and the complex successfully inhibited melanoma lung metastasis. This biodegradable and effective siRNA delivery vector, siRNA ternary complex, could be available for clinical trials.

Molecular glues for protein-protein interactions: Progressing toward a new dream.

The modulation of protein-protein interactions with small molecules is one of the most rapidly developing areas in drug discovery. In this review, we discuss advances over the past decade (2014-2023) focusing on molecular glues (MGs)-monovalent small molecules that induce proximity, either by stabilizing native interactions or by inducing neomorphic interactions. We include both serendipitous and rational discoveries and describe the different approaches that were used to identify them. We classify the compounds in three main categories: degradative MGs, non-degradative MGs or PPI stabilizers, and MGs that induce self-association. Diverse, illustrative examples with structural data are described in detail, emphasizing the elements of molecular recognition and cooperative binding at the interface that are fundamental for a MG mechanism of action.

Frizzleds act as dynamic pharmacological entities.

The Frizzled family of transmembrane receptors (FZD1-10) belongs to the class F of G protein-coupled receptors (GPCRs). FZDs bind to and are activated by Wingless/Int1 (WNT) proteins. The WNT/FZD signaling system regulates crucial aspects of developmental biology and stem-cell regulation. Dysregulation of WNT/FZD communication can lead to developmental defects and diseases such as cancer and fibrosis. Recent insight into the activation mechanisms of FZDs has underlined that protein dynamics and conserved microswitches are essential for FZD-mediated information flow and build the basis for targeting these receptors pharmacologically. In this review, we summarize recent advances in our understanding of FZD activation, and how novel concepts merge and collide with existing dogmas in the field.

The mechanisms of ·OH formation in MnO2 and oxalate system: Implication for ATZ removal.

Manganese oxides (MnO2) are commonly prevalent in groundwater, sediment and soil. In this study, we found that oxalate (H2C2O4) dissolved MnO2, leading to the formation of Mn(II)/(III), CO2(aq) and reactive oxygen species (·CO2-/O2·-/H2O2/·OH). Notably, CO2(aq) played a crucial role in ·OH formation, contributing to the degradation of atrazine (ATZ). To elucidate underneath mechanisms, a series of reactions with different gas-liquid ratios (GLR) were conducted. At the GLR of 0.3, 3.76, and + ∞ 79.4 %, 5.32 %, and 5.28 % of ATZ were eliminated, in which the cumulative ·OH concentration was 39.6 µM, 8.11 µM, and 7.39 µM and the cumulative CO2(aq) concentration was 11.2 mM, 4.7 mM, and 2.8 mM, respectively. The proposed reaction pathway was that CO2(aq) participated in the formation of a ternary complex [C2O4-Mn(II)-HCO4·3 H2O]-, which converted to a transition state (TS) as [C2O4-Mn(II)-CO3-OH·3 H2O]-, then decomposed to a complex radical [C2O4-Mn(II)-CO3·3 H2O]·- and ·OH after electron transfer within TS. It was novel to discover the role of CO2(aq) for ·OH yielding during MnO2 dissolution by H2C2O4. This finding helps revealing the overlooked processes that CO2(aq) influenced the fate of ATZ or other organic compounds in environment and providing us ideas for new technique development in contaminant remediation. ENVIRONMENTAL IMPLICATION: Manganese oxides and oxalate are common in soil, sediment and water. Their interactions could induce the formation of Mn(II)/(III), CO2(aq) and ·CO2-/O2·-/H2O2. This study found that atrazine could be effectively removed due to ·OH radicals under condition of high CO2(aq) concentration. The concentrations of Mn (0.0002-8.34 mg·L-1) and CO2(aq) (15-40 mg·L-1) were high in groundwater, and the surface water or rainfall seeps into groundwater and bring organic acids, which might promote the ·OH formation. The results might explain the missing steps of herbicides transformation in these environments and be helpful in developing new techniques in remediation in future.

Modification mechanism of soybean protein isolate-soluble soy polysaccharide complex by EGCG through covalent and non-covalent interaction: Structural, interfacial, and functional properties.

Soybean protein isolate was modified with polysaccharides and polyphenols to prepare a natural emulsifier with antioxidant capacity. Physicochemical, structural, interfacial, and functional properties of SPI-SSPS complex were investigated after covalent and non-covalent interacted with EGCG. SPI-SSPS-EGCG ternary complex with low EGCG concentrations (0.0625 and 0.125 mg/mL) showed a significant increase in absolute potential value and a decrease in turbidity. EGCG destroyed the original rigid structure of SPI-SSPS complex, and the covalent complexes had an ordered structure, while the non-covalent interaction resulted in disordered. The ternary complex with high EGCG concentrations (0.25 and 0.5 mg/mL) exhibited stronger EGCG binding capacity and lower surface hydrophobicity, which in turn affected its interfacial properties. The EAI and ESI of SPI-SSPS-EGCG covalent complex increased significantly, while the non-covalent complex had a significant change in EAI but no significant change in ESI with increasing EGCG concentration. The ternary complex showed significantly enhanced antioxidant capacity. The SPI-SSPS-EGCG ternary complex, with excellent antioxidant capacity and emulsifying property, making it suitable for emulsion delivery systems.

Conformational landscape of soluble α-klotho revealed by cryogenic electron microscopy.

α-Klotho (KLA) is a type-1 membranous protein that can associate with fibroblast growth factor receptor (FGFR) to form co-receptor for FGF23. The ectodomain of unassociated KLA is shed as soluble KLA (sKLA) to exert FGFR/FGF23-independent pleiotropic functions. The previously determined X-ray crystal structure of the extracellular region of sKLA in complex with FGF23 and FGFR1c suggests that sKLA functions solely as an on-demand coreceptor for FGF23. To understand the FGFR/FGF23-independent pleiotropic functions of sKLA, we investigated biophysical properties and structure of apo-sKLA. Mass photometry revealed that sKLA can form a stable structure with FGFR and/or FGF23 as well as sKLA dimer in solution. Single particle cryogenic electron microscopy (cryo-EM) supported the dimeric structure of sKLA. Cryo-EM further revealed a 3.3Å resolution structure of apo-sKLA that overlays well with its counterpart in the ternary complex with several distinct features. Compared to the ternary complex, the KL2 domain of apo-sKLA is more flexible. 3D variability analysis revealed that apo-sKLA adopts conformations with different KL1-KL2 interdomain bending and rotational angles. The potential multiple forms and shapes of sKLA support its role as FGFR-independent hormone with pleiotropic functions. A comprehensive understanding of the sKLA conformational landscape will provide the foundation for developing klotho-related therapies for diseases.

Trace Cu(II) as an Efficient Electron-Transfer Shuttle for Reductive Deiodination of Refractory Iodinated Contrast Media Compounds.

Iodinated contrast media compounds (ICMs) are intensively applied in medical diagnostic radiology and have received wide environmental concerns due to formation potential of iodinated disinfection byproducts. Conventional water/wastewater treatment processes cannot effectively remove ICMs; reducing their total organic iodine concentration is even more difficult. The source control or elimination of ICMs thus becomes necessary. We report here that the refractory ICMs (5 µM) can be efficiently deiodinated by ascorbate/ascorbic acid (AA) (200 µM) coupled with a trace amount of Cu(II) (5 µM) through catalytic reduction but not oxidation, contrary to the conventional concept of AA/Cu(II) coupling, which produces reactive oxygen species. Taking diatrizoate (DTZ, a refractory ICM) as an example, the coupling completely deiodinated DTZ without destroying its molecular structure. High-performance liquid chromatography inductively coupled plasma mass spectrometry analysis revealed that ternary complexes form between Cu(II), ascorbate, and the anilide moiety of DTZ. Cu(II) in the ternary complex works as an efficient electron-transfer shuttle to convey electrons from ascorbate to the target compound, inducing sequential and complete deiodination. Both DTZ and the nonionic ICMs can be effectively deiodinated even in human urine. Thus, AA coupled with trace Cu(II) could be potentially useful for the source elimination of organic iodine of ICMs.