Binary Complex Research Articles

Antitumor potential of platinum(II) complexes of selenium donor ligands.

Platinum(II) coordination compounds are widely applied in clinics as anticancer drugs. In this review, we provide a summary of the reports on cytotoxic properties of platinum(II) complexes of selenium donor ligands along with a brief description of their structural features. It has been observed that the platinum(II) complexes of selenones and selenoethers display reasonable antitumor properties and in some cases their cytotoxic activity is greater than cisplatin. The complexes containing NH3 ligands along with selenones were found to exhibit better cytotoxicity compared to the binary Pt-selenone complexes. The mechanistic insights showed that these complexes exert antitumor activity through reactive oxygen species (ROS) generation and induction of apoptosis. The platinum-selenoether coordination compounds can self-assemble into spherical aggregates capable of self-delivery. The self-assembled Pt-selenium aggregates induce cell apoptosis via ROS, which leads to high selectivity between cancer cells and normal cells in cytotoxicity assays.

Conformations and non-covalent interactions of cyclohexyl isothiocyanate and its water complex

The conformational equilibria of cyclohexyl isothiocyanate and its complex with water were studied by microwave spectroscopy combined with theoretical calculations. For the bare cyclohexyl isothiocyanate, the rotational spectra of the two conformers, namely a-trans and e-trans, were measured and analyzed. Additional mono-substituted isotopologues of two 13C and one 34S of a-trans and e-trans were also measured, which result in an accurate structural determination of the two conformers. For the binary complex with water, the rotational spectra were measured for the monohydrates of both a-trans and e-trans conformers. Water isotopologues were also detected for the monohydrates. All the measured rotational spectra show 14N quadrupole coupling hyperfine structures. Water molecule prefers linking to the isothiocyanate group through a OwH⋯S hydrogen bond and forming two CH⋯Ow hydrogen bonds with the oxygen atom of water acting as a proton acceptor. The non-covalent bonding features of the monohydrates were revealed by natural bond orbital analysis and symmetry-adapted perturbation theory analysis.

Structures of apo Cas12a and its complex with crRNA and DNA reveal the dynamics of ternary complex formation and target DNA cleavage.

Cas12a is a programmable nuclease for adaptive immunity against invading nucleic acids in CRISPR-Cas systems. Here, we report the crystal structures of apo Cas12a from Lachnospiraceae bacterium MA2020 (Lb2) and the Lb2Cas12a+crRNA complex, as well as the cryo-EM structure and functional studies of the Lb2Cas12a+crRNA+DNA complex. We demonstrate that apo Lb2Cas12a assumes a unique, elongated conformation, whereas the Lb2Cas12a+crRNA binary complex exhibits a compact conformation that subsequently rearranges to a semi-open conformation in the Lb2Cas12a+crRNA+DNA ternary complex. Notably, in solution, apo Lb2Cas12a is dynamic and can exist in both elongated and compact forms. Residues from Met493 to Leu523 of the WED domain undergo major conformational changes to facilitate the required structural rearrangements. The REC lobe of Lb2Cas12a rotates 103° concomitant with rearrangement of the hinge region close to the WED and RuvC II domains to position the RNA-DNA duplex near the catalytic site. Our findings provide insight into crRNA recognition and the mechanism of target DNA cleavage.

Structural Basis for Substrate Binding and Catalytic Mechanism of the Key Enzyme VioD in the Violacein Synthesis Pathway.

Violacein is a pigment synthesized by gram-negative bacteria with various biological activities such as antimicrobial, antiviral, and anticancer activities. VioD is a key oxygenase converting protodeoxyviolaceinic acid (PDVA) to protoviolaceinic acid in violacein biosynthesis. To elucidate the catalytic mechanism of VioD, here, we resolved two crystal structures of VioD, a binary complex structure containing VioD and a FAD and a ternary complex structure composed of VioD, a FAD and a 2-ethyl-1-hexanol (EHN). Structural analysis revealed a deep funnel like binding pocket with wide entrance, this pocket is positively charged. The EHN is located at the deep bottom of the binding pocket near isoalloxazine ring. Further docking simulation help us to propose the mechanism of the hydroxylation of the substrate catalyzed by VioD. Bioinformatic analysis suggested and emphasized the importance of the conserved residues involved in substrate binding. Our results provide a structural basis for the catalytic mechanism of VioD. This article is protected by copyright. All rights reserved.

Edmond Fischer's kinase legacy: History of the protein kinase inhibitor and protein kinaseA.

Although Fischer's extraordinary career came to focus mostly on the protein phosphatases, after his co-discovery of Phosphorylase Kinase with Ed Krebs he was clearly intrigued not only by cAMP-dependent protein kinase (PKA), but also by the heat-stable, high-affinity protein kinase inhibitor (PKI). PKI is an intrinsically disordered protein that contains at its N-terminus a pseudo-substrate motif that binds synergistically and with high-affinity to the PKA catalytic (C) subunit. The sequencing and characterization of this inhibitor peptide (IP20) were validated by the structure of the PKA C-subunit solved first as a binary complex with IP20 and then as a ternary complex with ATP and two magnesium ions. A second motif, nuclear export signal (NES), was later discovered in PKI. Both motifs correspond to amphipathic helices that convey high-affinity binding. The dynamic features of full-length PKI, recently captured by NMR, confirmed that the IP20 motif becomes dynamically and sequentially ordered only in the presence of the C-subunit. The type I PKA regulatory (R) subunits also contain a pseudo-substrate ATPMg2-dependent high-affinity inhibitor sequence. PKI and PKA, especially the Cβ subunit, are highly expressed in the brain, and PKI expression is also cell cycle-dependent. In addition, PKI is now linked to several cancers. The full biological importance of PKI and PKA signaling in the brain, and their importance in cancer thus remains to be elucidated.

ORP9 and ORP10 form a heterocomplex to transfer phosphatidylinositol 4-phosphate at ER-TGN contact sites.

Oxysterol-binding protein (OSBP) and its related proteins (ORPs) are a family of lipid transfer proteins (LTPs) that mediate non-vesicular lipid transport. ORP9 and ORP10, members of the OSBP/ORPs family, are located at the endoplasmic reticulum (ER)-trans-Golgi network (TGN) membrane contact sites (MCSs). It remained unclear how they mediate lipid transport. In this work, we discovered that ORP9 and ORP10 form a binary complex through intermolecular coiled-coil (CC) domain-CC domain interaction. The PH domains of ORP9 and ORP10 specially interact with phosphatidylinositol 4-phosphate (PI4P), mediating the TGN targeting. The ORP9-ORP10 complex plays a critical role in regulating PI4P levels at the TGN. Using in vitro reconstitution assays, we observed that while full-length ORP9 efficiently transferred PI4P between two apposed membranes, the lipid transfer kinetics was further accelerated by ORP10. Interestingly, our data showed that the PH domains of ORP9 and ORP10 participate in membrane tethering simultaneously, whereas ORDs of both ORP9 and ORP10 are required for lipid transport. Furthermore, our data showed that the depletion of ORP9 and ORP10 led to increased vesicle transport to the plasma membrane (PM). These findings demonstrate that ORP9 and ORP10 form a binary complex through the CC domains, maintaining PI4P homeostasis at ER-TGN MCSs and regulating vesicle trafficking.

Precise analysis of thyroxine enantiomers in pharmaceutical formulation by mobility difference based on cyclodextrin

Identification and determination of chiral pharmaceutical residues is still a challenging analytical puzzle. In this work, a simple, rapid, and effective method for chiral D/L-tetraiodothyronine (T4) separation and quantitative was developed based on host–guest recognition using ion mobility spectrometry-mass spectrometry (IMS-MS). The D/L-T4 enantiomers were mobility separated by their diastereomeric complexes through mixing with cyclodextrin (CD) and metal ions. D/L-T4 was first separated by complexing with host molecule (α-, β-, γ-CD), observing weak peak-to-peak resolution (Rp-p) by the formed binary complex [CD + D/L-T4-H]+, and the Rp-p decreased with the CD size increasing. However, the separation effect of D/L-T4 was much improved with the addition of divalent metal ions (G2+) by the formed ternary complex [CD + D/L-T4 + G]2+. In comparison, α-CD related complexes can possess the best separation effect for D/L-T4 in most cases. Considering the high selectivity, non-toxic, and chemically stable of β-CD, [β-CD + D/L-T4 + Ca]2+ was selected for D/L-T4 analysis (RP-P = 0.764). Whereafter, chemical theoretical conformations for [β-CD + D/L-T4 + H]+ and [β-CD + D/L-T4 + Ca]2+ were optimized, discovering similar micro-interaction modes between [β-CD + D-T4 + H]+ and [β-CD + L-T4 + H]+; while with the addition of Ca2+, significantly different interaction modes were observed between [β-CD + D-T4 + Ca]2+ and [β-CD + L-T4 + Ca]2+. And theoretical collision cross section (CCS) trends for the complexes were consistent with that of the experimental results. Additionally, calibration curves were linear within 1.00 to 104 ng mL−1 with coefficient (R2 > 0.99), gaining the limit of detection (LODs) calculation of 0.11 ng mL−1, and the detection range between D-T4 and L-T4 of 45.6:1 to 1:59.8. Finally, the method was applied for D/L-T4 detection in Levothyroxine tablets, the detection content has good consistency on drug labeling. Because the proposed method exhibited good analytical performance in terms of speed, selectivity, sensitivity, and reproducibility of the measurements, that can be a promising strategy for effective D/L-T4 detection in pharmaceutical industries or other practical samples.

Regulation of cargo exocytosis by a Reps1-Ralbp1-RalA module.

Surface levels of membrane proteins are determined by a dynamic balance between exocytosis-mediated surface delivery and endocytosis-dependent retrieval from the cell surface. Imbalances in surface protein levels perturb surface protein homeostasis and cause major forms of human disease such as type 2 diabetes and neurological disorders. Here, we found a Reps1-Ralbp1-RalA module in the exocytic pathway broadly regulating surface protein levels. Reps1 and Ralbp1 form a binary complex that recognizes RalA, a vesicle-bound small guanosine triphosphatases (GTPase) promoting exocytosis through interacting with the exocyst complex. RalA binding results in Reps1 release and formation of a Ralbp1-RalA binary complex. Ralbp1 selectively recognizes GTP-bound RalA but is not a RalA effector. Instead, Ralbp1 binding maintains RalA in an active GTP-bound state. These studies uncovered a segment in the exocytic pathway and, more broadly, revealed a previously unrecognized regulatory mechanism for small GTPases, GTP state stabilization.

Engineering a novel rhodamine-based fluorescent probe using host-guest interactions for reversible, selective, and sensitive detection of herbicide paraquat

Although paraquat (PQ) is one of the most extensively used herbicides worldwide, it is hazardous to the environment and the public due to its extreme toxicity towards non-target organisms especially animals and humans. Therefore, developing a detection system for selectively and sensitively detecting PQ remains a great challenge. This report demonstrates the design of a novel highly fluorescent rhodamine-based probe fabricated by rationally inserting adamantane-modified rhodamine into a host cucurbit[7]uril (CB[7]) with a host-guest interaction induced spirocyclic ring-opening process. This host-guest binary complex (1&CB[7]) had a stronger fluorescence intensity than rhodamine B, which was exploited for PQ detection. Notably, complex 1&CB[7] displayed high sensitivity toward PQ, with an excellent nanomolar limit of detection of 10 nM. It was determined that this unique detection mechanism involved a process of self-assembly, disassembly, and reassembly, thereby providing a reversible fluorescence response to PQ. With such merits, we successfully quantified and mapped PQ in actual water samples, live cells, and various agricultural products. We anticipated that the constructed complex 1&CB[7] could acts as a potential alternative for PQ tracking in various applications, highlighting a new paradigm for engineering host-guest probes with spirocyclic ring-opening process for analyte detection.

Complexation between the Antioxidant Pterostilbene and Derivatized Cyclodextrins in the Solid State and in Aqueous Solution.

Inadequate aqueous solubilities of bioactive compounds hinder their ability to be developed for medicinal applications. The potent antioxidant pterostilbene (PTB) is a case in point. The aim of this study was to use a series of modified water-soluble cyclodextrins (CDs), namely, hydroxypropyl β-CD (HPβCD), dimethylated β-CD (DIMEB), randomly methylated β-CD (RAMEB), and sulfobutyl ether β-CD sodium salt (SBECD) to prepare inclusion complexes of PTB via various solid, semi-solid, and solution-based treatments. Putative CD-PTB products generated by solid-state co-grinding, kneading, irradiation with microwaves, and the evaporative treatment of CD-PTB solutions were considered to have potential for future applications. Primary analytical methods for examining CD-PTB products included differential scanning calorimetry and Fourier transform infrared spectroscopy to detect the occurrence of binary complex formation. Phase solubility analysis was used to probe CD-PTB complexation in an aqueous solution. Complexation was evident in both the solid-state and in solution. Complex association constants (K1:1) in an aqueous solution spanned the approximate range of 15,000 to 55,000 M-1; the values increased with the CDs in the order HPβCD < DIMEB < RAMEB < SBECD. Significant PTB solubility enhancement factors were recorded at 100 mM CD concentrations, the most accurately determined values being in the range 700-fold to 1250-fold.

NEW PROMISING AGENTS FOR PHOTODYNAMIC THERAPY

The problem of oncological diseases has acquired a social and medical character. In this regard, the rapid development of new drugs and methods, as well as modification of existing methods in order to increase their effectiveness, is currently required. The paper aims to develop and study two promising agents for photodynamic therapy. We discussed a new binary complex based on the widely used photosensitizer chlorin E6 with europium, which can solve one of the main disadvantages of drugs for photodynamic therapy: it can be used to treat deeper tumors and act more effectively by transferring luminescence energy between europium and chlorin E6. Our other agent was copper-containing chlorophyllin. Chlorophyllin is a natural component, which is always promising in the development of new drugs. The optical and fluorescent parameters of the agents were determined using physicochemical methods; their ability to generate reactive oxygen species in the presence of two fluorescent probes was studied: 2ʹ, 7ʹ-dichlorodihydrofluorescein and dihydrofluorescein. Thus, we showed the effectiveness of chlorophyllin in comparison with chlorin E6 by 1.92 times at 50 μg / ml E6 and showed the effectiveness of the binary complex even at the minimum concentration of 10 μg / ml. Further research in cell cultures will develop the ways of the possible use of new substances as photosensitizers.

Dynamics of ternary complex formation of an IDP and two binding partners and their allosteric effects studied by three-color single-molecule FRET.

Intrinsically disordered proteins (IDPs) are highly flexible and can bind multiple binding partners by folding into different structures. This promiscuity of IDPs is especially important for their function as hub proteins in the formation protein-protein interaction networks, which is a key regulatory mechanism of complex biological processes such as gene transcription and signal transduction. IDPs often have more than one binding domain, suggesting they can interact with multiple binding partners simultaneously. Ternary complexes formed by an IDP and two binding partners have been observed by NMR, but it is difficult to obtain more detailed kinetic information from ensemble measurements. In this work, we used three-color single-molecule FRET to directly visualize the formation and dissociation of a ternary complex. We studied binding of an IDP (p53 transactivation domain (TAD)) and two competitive binding partners (TAZ2 of CBP and MDM2). By labeling TAD with Alexa 488 (donor) and Alexa 647 (acceptor 1) and TAZ2 with CF680R (acceptor 2), it is possible to detect the formation of not only two binary complexes (TAD-TAZ2 and TAD-MDM2) but also the ternary complex (TAD-MDM2-TAZ2), which can be distinguished by the fraction of acceptor 2 intensity. From the kinetic analysis, we found that the TAD-TAZ2 binary complex is the most stable and the ternary complex is formed only transiently. Interestingly, the presence of MDM2 induces more rapid dissociation of TAZ2 in the ternary complex whereas the dissociation rate of MDM2 is not affected by TAZ2. This asymmetric allosteric effect results in the more effective replacement of CBP on p53 with MDM2, which is a novel regulatory mechanism beyond the simple competition between multiple binding partners.

Biophysics of IL-22 receptor interaction with selected interleukins.

Atopic dermatitis (AD) is the most prevalent chronic inflammatory skin condition globally, and many patients do not respond adequately to currently available therapies [1]. Interleukin-22 receptor 1 (IL-22RA1) has been identified as a potential mediator of AD, but no treatments for this pathway are currently on the market [2]. In this work, we aim to elucidate the biophysics of the binary and ternary IL-22RA1 complex formation with interleukins by combining surface plasmon resonance requiring protein immobilization and in-solution techniques such as small angle X-ray scattering and circular dichroism. In addition, structural characterization of the ternary complexes by high-resolution cryogenic electron microscopy is ongoing and will be used to provide Angstrom resolution structural data about the complexes. This will be coupled to molecular dynamics simulations, providing a comprehensive overview of the formation of IL-22RA1 signaling complexes resulting in knowledge on the domains involved in the signaling process. This will result in new and novel knowledge on the interaction with these interleukins and IL22R1 and provide insights into the role of these interleukins and potential approaches to block their interaction by an antibody approach.

Breastfeeding status and determinants of current breastfeeding of Syrian refugee children in Turkey

BackgroundTurkey hosts the highest number of refugees in the World including 65% of Syrian refugees who reside in Turkey. Mothers and children were the most negatively affected among the Syrian refugees who had to migrate from their countries as a result of the civil war in Syria. One of the most important issues in terms of child health is breastfeeding. Breastfeeding in migrants should be promoted worldwide to mitigate infant mortality and diseases. The aim of this study is to examine the association between breastfeeding status in Syrian refugee children under two years and socio-demographic characteristics of Syrian refugee mothers with further analysis of Turkey Demographic and Health Survey-Syrian Migrant-2018 (TDHS-SM-2018) data. MethodsThe data source is the TDHS-SM-2018. Data for the last-born children with a gestational duration greater than 32 weeks from the mothers’ singleton pregnancy, aged less than two years old and living with the mother were included (unweighted n = 744). The dependent variable was the breastfeeding status (breastfeeding in the last 24 h during the study period) in children under two years. Complex sample logistic regression evaluated the associations.ResultsThe percentage of breastfeeding in children under the age of two years was found to be 62.4%, and the total median breastfeeding duration was 14.6 months. Univariate analysis showed that the earlier mothers immigrated to Turkey, the higher the current breastfeeding rate. Breastfeeding rates were found to be higher among people living in the South and East regions (65.2% and 65.1% respectively). Multivariable binary complex sample logistic regression revealed that breastfeeding status at the study period was associated with long preceding birth interval; delivery in a public hospital; absence of prelacteal feeding; being non-pregnant; and the region and age of the child. No relationship for current breastfeeding was found with maternal activities, maternal life satisfaction, financial satisfaction, and educational status.ConclusionCurrent breastfeeding in our sample was more likely among mothers with a longer birth interval who avoided prelacteal feeding. The Baby-Friendly approach and family planning services should be integrated into refugee health centers.

Understanding mucus modulation behavior of chitosan oligomers and dextran sulfate combining light scattering and calorimetric observations

This study reports the fundamental understanding of mucus-modulatory strategies combining charged biopolymers with distinct molecular weights and surface charges. Here, key biophysical evidence supports that low-molecular-weight (Mw) polycation chitosan oligosaccharides (COSs) and high-Mw polyanion dextran sulfate (DS) exhibit distinct thermodynamic signatures upon interaction with mucin (MUC), the main protein of mucus. While the COS → MUC microcalorimetric titrations released ~14 kcal/mol and ~60 kcal/mol, the DS → MUC titrations released ~1200 and ~1450 kcal/mol at pH of 4.5 and 6.8, respectively. The MPT-2 titrations of COS → MUC and DS → MUC indicated a greater zeta potential variation at pH = 4.5 (relative variation = 815 % and 351 %, respectively) than at pH = 6.8 (relative variation = 282 % and 136 %, respectively). Further, the resultant binary (COS-MUC) and ternary (COS-DS-MUC) complexes showed opposite behavior (aggregation and charge inversion events) according to the pH environment. Most importantly, the results indicate that electrostatics could not be the driving force that governs COS-MUC interactions. To account for this finding, we proposed a two-level abstraction model. Macro features emerge collectively from individual interactions occurring at the molecular level. Therefore, to understand the outcomes of mucus modulatory strategy based on charged biopolymers it is necessary to integrate both visions into the same picture.

Towards a structurally resolved human protein interaction network

Cellular functions are governed by molecular machines that assemble through protein-protein interactions. Their atomic details are critical to studying their molecular mechanisms. However, fewer than 5% of hundreds of thousands of human protein interactions have been structurally characterized. Here we test the potential and limitations of recent progress in deep-learning methods using AlphaFold2 to predict structures for 65,484 human protein interactions. We show that experiments can orthogonally confirm higher-confidence models. We identify 3,137 high-confidence models, of which 1,371 have no homology to a known structure. We identify interface residues harboring disease mutations, suggesting potential mechanisms for pathogenic variants. Groups of interface phosphorylation sites show patterns of co-regulation across conditions, suggestive of coordinated tuning of multiple protein interactions as signaling responses. Finally, we provide examples of how the predicted binary complexes can be used to build larger assemblies helping to expand our understanding of human cell biology.

In-Vitro Diagnostic Reagent Evaluation of Commercially Available Cardiac Troponin I Assay Kits Using H/D Exchange Mass Spectrometry for Antibody-Epitope Mapping.

Cardiac troponin I (cTnI) is the biomarker of choice and considered a gold standard for the diagnosis of acute myocardial infarction. However, the quantitative results of cTnI assay kits from different manufacturers are not comparable. Based on the H/D exchange mass spectrometry (HDX-MS) workflow, we developed an in-vitro diagnostic reagent antibody evaluation strategy to analyze the interactions of epitopes and antibody cocktails─(R195, F12, S13) and (D1, D2, pAb2). The HDX results indicate that the quantitative result bias of the different reagents originates from the ability of antibodies to recognize various cTnI complex forms, such as free cTnI, hydrolyzed cTnI, and cTnI combined with cTnT or TnC as binary or ternary complexes (cTnIC, cTnTIC), in blood based on different epitopes. The data obtained from the peptide HDX of interest after treatment with various antibody cocktails clearly indicated epitope specificity. The consistency of quantitative results can be improved by a thorough investigation into the epitopes recognized by the antibodies of various diagnostic kits, which will lead to the standardization of cTnI diagnosis.

Spectrofluorimetric assay of amlodipine and perindopril in their raw materials, pharmaceutical formulations and spiked human plasma through the formation of complexes with Eosin Y

For quantitation of amlodipine (AML) and perindopril (PER) in their authentic, pharmaceutical formulations and spiked human plasma, a simple, sensitive, validated and inexpensive spectrofluorimetric method has been developed. The proposed method is developed to be based on quantitative quenching effect of two antihypertensive drugs on Eosin Y's native fluorescence which was achieved by developing binary complexes between each of the cited drugs in an acidic environment using acetate buffer (pH 4.4) with Eosin Y. Fluorescence quenching was recorded at 544 nm after excitation at 425 nm. For AML and PER, calibration curves were obtained over the range of 0.3–3.0 µg/mL and 0.2–2.0 µg/mL, respectively, with correlation coefficients of 0.9993 and 0.9995, respectively. The developed method was validated according to ICH guidelines. The proposed spectrofluorimetric method is regarded new and sensitive. As a result, the proposed method might be used to estimate the quality of the cited drugs in their pharmaceutical formulations and biological fluid.

Structural mechanisms of calmodulin activation of Shigella effector OspC3 to ADP-riboxanate caspase-4/11 and block pyroptosis.

The caspase-4/11-GSDMD pyroptosis axis recognizes cytosolic lipopolysaccharide for antibacterial defenses. Shigella flexneri employs an OspC3 effector to block pyroptosis by catalyzing NAD+-dependent arginine ADP-riboxanation of caspase-4/11. Here, we identify Ca2+-free calmodulin (CaM) that binds and stimulates OspC3 ADP-riboxanase activity. Crystal structures of OspC3-CaM and OspC3-caspase-4 binary complexes reveal unique CaM binding to an OspC3 N-terminal domain featuring an ADP-ribosyltransferase-like fold and specific recognition of caspase-4 by an OspC3 ankryin repeat domain, respectively. CaM-OspC3-caspase-4 ternary complex structures show that NAD+ binding reorganizes the catalytic pocket, in which D231 and D177 activate the substrate arginine for initial ADP-ribosylation and ribosyl 2'-OH in the ADP-ribosylated arginine, respectively, for subsequent deamination. We also determine structures of unmodified and OspC3-ADP-riboxanated caspase-4. Mechanisms derived from this series of structures covering the entire process of OspC3 action are supported by biochemical analyses in vitro and functional validation in S. flexneri-infected mice.

Improved thermal stability of roselle anthocyanin by co-pigmented with oxalic acid: Preparation, characterization and enhancement mechanism

The enhancement effects of co-pigmentation on thermal stability of roselle anthocyanin extract (RAE) were investigated. The introduction of organic acids maintained color stability of RAE, and RAE co-pigmented with oxalic acid (OA) presented less color fading rates (19.46 ± 0.33 %) and higher redness (41.33 ± 3.51). Subsequently, suitable co-pigmentation concentration (OA:RAE = 1:2) was obtained regarding with lower ΔE (48.70 ± 2.36). Then, improvement behaviors of co-pigmentation on OA-RAE were evaluated. Results demonstrated that OA-RAE exhibited better thermal stability, as manifested by larger retention rates and more favorable thermal degradation kinetic parameters. Furthermore, both molecular docking simulation and experimental structural characterization revealed that hydrogen bonds and other non-covalent bonds made up the main parts of molecular interactions, leading to formation of stable binary complex. As a result, the aromatic ring of RAE was protected. In conclusion, the co-pigmentation of RAE via introduction of OA was effective in stability enhancement due to the generation of molecular bindings.