Insight into the efficient loading and enhanced activity of enzymes immobilized on functionalized UiO-66.

Enzyme immobilization is an effective strategy for achieving efficient and sustainable enzyme catalysis. As a kind of promising enzyme-loading materials, the systematic research on zirconium based metal organic frameworks (Zr-MOFs) about immobilization performance at molecular level is still in its initial stage. In this work, UiO-66 was functionalized with various groups (-H, -NH2, -COOH, -OH, -2OH) for the immobilization of cytochrome c (Cyt c) and antioxidant enzyme catalase (CAT). Then the effects of surface-functionalized UiO-66 derivatives on the loading efficiency, enzyme stability and catalysis kinetics were systematically investigated. In addition, the affinity constants of Cyt c and CAT towards UiO-66-series MOFs carriers were also compared. The results have shown that hydroxyl group functionalized UiO-66 represents the highest enzyme loading capacity, enhanced activity and improved stability for Cyt c and CAT possibly due to high surface area and suitable microenvironments as well as enhanced affinity towards the enzymes provided by the introduction of a single hydroxyl group. Our research would foresee immense potential of MOFs in engineering biocatalysts.

Fabrication of a Terbium-Functionalized Cadmium Organic Framework with Proper Energy Levels as a Ratiometric Probe of an Anthrax Biomarker.

Anthrax bacillus is a very dangerous zoonotic pathogen that seriously endangers public health. Rapid and accurate qualitative and quantitative detection of its biomarkers, 2,6-dipicolinic acid (DPA), is crucial for the prevention and treatment of this pathogenic bacterium. In this work, a novel Cd-based MOF (TTCA-Cd) has been synthesized from a polycarboxylate ligand, [1,1':2',1″-terphenyl]-4,4',4″,5'-tetracarboxylic acid (H4TTCA), and further doped with Tb(III), forming a dual-emission lanthanide-functionalized MOF hybrid (TTCA-Cd@Tb). TTCA-Cd@Tb can be developed as a high-performance ratiometric fluorescent sensor toward DPA with a very low detection limit of 7.14 nM and high selectivity in a wide detection range of 0-200 µM, demonstrating a big advancement and providing a new option for the detection of DPA.

Four three-dimensional rare earth metal - organic framework fluorescent sensor for efficient detection of gentamicin sulfate and Fe3.

Excessive use of gentamicin sulfate can cause severe nephrotoxicity and ototoxicity, abnormal levels of Fe3+ intake can also cause serious damage to body. Therefore, establishing a fast and accurate detection method for the above-mentioned substances is of great significance. However, traditional detection methods such as high-performance liquid chromatography still have certain problems such as high cost and complex operation. Fluorescent MOFs are favored by analysts due to their high specific surface area, high porosity, adjustable pore size, and good stability. In this paper, we have synthesized four rare earth MOFs based on the pyridinecarboxylic acid ligand (H2L), which are [Eu(L)1/2H2O]n, [Gd(L)1/2H2O]n, [Sm(L)1/2H2O]n, [Y(L)3/2H2O·DMF]n. The structures of four MOFs were confirmed by single crystal X-ray diffraction, which proved that MOF-1, MOF-2 and MOF-3 were isostructural, and all the four MOFs were three-dimensional structures. In the fluorescence test, gentamicin sulfate and Fe3+ can cause significant fluorescence quenching of MOF-1 and MOF-4 respectively, and show good selectivity and anti-interference performance, as well as low detection limit and wide detection range. This work may provide a possibility for the detection of gentamicin sulfate and iron ions in complex environments.

Blue-Emitting Zero-Dimensional Inorganic-Organic Hybrids Constructed from Beta-Diketonate Ligands and Bulky Organic Cations.

Two zero-dimensional inorganic-organic hybrids, namely, [C4mim][Cd(TCDPPA)3] (1) and [C4mpy][Cd(TCDPPA)3] (2), where (TCDPPA)- = 2,2,2-trichloro-N-(di(pyrrolidin-1-yl)phosphoryl)acetamide, (C4mim)+ = 1-butyl-3-methylimidazolium, and (C4mpy)+ = 1-butyl-4-methylpyridinium, have been synthesized via metathesis reactions and characterized systematically. These ionic cadmium-containing inorganic-organic hybrid compounds are assembled from a bulky organic cation and a complex anion constructed from the chelation of three TCDPPA ligands to one cadmium ion. These compounds possess wide band gaps and emit in the deep-blue region intensely with a quantum yield as high as 34.04%. The success of this work provides a new method for the design and fabrication of high-efficiency blue-emitting materials.

Functionalized Zirconium Organic Frameworks as Fluorescent Probes for the Detection of Tetracyclines in Water and Pork.

The overuse of tetracyclines (TCs) in livestock breeding may cause a series of health and environmental problems. It is necessary to develop more accurate, convenient, and rapid sensing methods toward TCs, but it is still very challenging. In this work, three isostructural zirconium organic frameworks (Zr-MOFs) have been investigated as probes for the fluorescent sensing of TCs in water. By varying the functional group at the central benzene core, their sensing performances toward TCs can be modified. Under optimized conditions, the limit of detection can be as low as 0.08 nM in a wide detection range of 0-147 µM with high sensitivity and selectivity. These Zr-MOFs can also be applied in the detection of TCs in real pork samples with satisfying reliabilities and correctness. This work provides a new method for the design and optimization of fluorescent sensors toward TCs.

Lanthanide-uranyl phosphonates constructed from diethyl ((phenylsulfonyl)methyl)phosphonate.

Lanthanide-uranyl phosphonates possess intriguing crystal structures and huge application prospects, but the construction of these materials remains challenging. In this work, we demonstrate that new uranyl and lanthanide-uranyl sulfonylphosphonates with elegant crystal structures and photophysical properties can be assembled hydrothermally by employing the heterofunctional diethyl phosphonate (diethyl ((phenylsulfonyl)methyl)phosphonate, Et2L) as a precursor ligand, which provides an effective strategy for the construction of lanthanide-uranyl phosphonates.

Ready Access to Anhydrous Anionic Lanthanide Acetates by Using Imidazolium Acetate Ionic Liquids as the Reaction Medium.

Access to lanthanide acetate coordination compounds is challenged by the tendency of lanthanides to coordinate water and the plethora of acetate coordination modes. A straightforward, reproducible synthetic procedure by treating lanthanide chloride hydrates with defined ratios of the ionic liquid (IL) 1-ethyl-3-methylimidazolium acetate ([C2 mim][OAc]) has been developed. This reaction pathway leads to two isostructural crystalline anhydrous coordination complexes, the polymeric [C2 mim]n [{Ln2 (OAc)7 }n ] and the dimeric [C2 mim]2 [Ln2 (OAc)8 ], based on the ion size and the ratio of IL used. A reaction with an IL : Ln-salt ratio of 5 : 1, where Ln=Nd, Sm, and Gd, led exclusively to the polymer, whilst for the heaviest lanthanides (Dy-Lu) the dimer was observed. Reaction with Eu and Tb resulted in a mixture of both polymeric and dimeric forms. When the amount of IL and/or the size of the cation was increased, the reaction led to only the dimeric compound for all the lanthanide series. Crystallographic analyses of the resulting salts revealed three different types of metal-acetate coordination modes where η2 µκ2 is the most represented in both structure types.

Probing the toxic interactions between bisphenol A and glutathione S-transferase Phi8 from Arabidopsis thaliana.

As primary polymer material in industrial products, bisphenol A (BPA) has become one of the most productive chemicals. Excluding its endocrine-disrupting property, BPA can also produce excessive reactive oxygen species (ROS). Nevertheless, the underlying toxic mechanisms of BPA-induced oxidative damages to plants are still unknown. In this work, glutathione S-transferase Phi8 was used as biomarker to evaluate the hazardous oxidative effects of BPA at the molecular level. Firstly, the intrinsic fluorescence of AtGSTF8 was statically quenched along with complex formation and structural and conformational changes, which led to the loosening and unfolding of the framework of AtGSTF8 as well as the increase of hydrophilicity around Trp residues. Then a single binding site was predicted for AtGSTF8 towards BPA and the complex formation was predominantly driven by hydrophobic interactions owing to the positive ΔH and ΔS. Besides, the predicted binding site of BPA was close to the H-site of AtGSTF8 which was surrounded by several hydrophobic amino acids based on the molecular docking results. The activity of glutathione S-transferase was declined and the plant growth was destroyed upon complex formation. The investigation of the binding mechanism of BPA with AtGSTF8 at molecular level would provide experimental assessments on toxicological effects of BPA on plants.

Speciation of Ionic Uranyl-Containing Complexes in in Situ Formed Dicyanonitrosomethanide-Based Ionic Liquids.

A series of ionic uranyl-containing complexes, namely [C2mim]2[UO2(ccnm)4] (1), [C4mim]2[UO2(ccnm)4] (2), [N1111]2[UO2(ccnm)4][H2O]2 (3), and [P2444]2[UO2(dcnm)2(ccnm)2] (4) [(ccnm)- = carbamoylcyanonitrosomethanide; dcnm = dicyanonitrosomethanide; (C2mim)+ = 1-ethyl-3-methylimidazolium; (C4mim)+ = 1-butyl-3-methylimidazolium; (N1111)+ = tetramethylammonium; (P2444)+ = tributyl(ethyl)phosphonium)], were isolated from in situ formed dcnm-based ionic liquids and characterized systematically. It was found that the dcnm anions transformed into ccnm anions during the reactions. These anions coordinate with the uranyl cations in chelate or terminal monodentate coordination mode, affording negative divalent complex anions which can combine with different organic cations and form ionic uranyl-containing complexes. Plenty of C-H···O, N-H···O, C-H···N, N-H···N, and H···H weak interactions are formed in the crystal structures. The transformation of cyano to amide groups contributes to the crystallinity and leads to higher melting points as well as the luminescence quenching of these compounds.

Variability of Uranyl Carboxylates from Rigid Terophenyldicarboxylic Acid Ligands.

Three uranyl carboxylates, namely, (UO2)(L1)(H2O)0.5 (1), [(UO2)(L2)(H2O)]·2H2O (2), and [(UO2)(L2)(H2O)]·(CH3CN) (3), were synthesized hydrothermally from 2',3',5',6'-tetramethyl-(1,1':4',1″-terphenyl)-4,4″-dicarboxylic acid (H2L1) and 2',5'-dimethyl-(1,1':4',1″-terphenyl)-3,3″-dicarboxylic acid (H2L2), which are all steric carboxylic acid ligands but vary with the carboxylic acid group position and methyl group number. It is found that compound 1 displays a three-dimensional 8-fold-interpenetrated net with channels running along the c direction. Compounds 2 and 3 are isostructural, and all display two-dimensional-layered crystal structures but contain different guest molecules. The photophysical measurements reveal that compounds 1 and 2, which contain disordered water molecules, are luminescence-quenched, whereas compound 3 containing acetonitrile molecules is luminescent.

Forcing Dicyanamide Coordination to f-Elements by Dissolution in Dicyanamide-Based Ionic Liquids.

A robust general route to lanthanide dicyanamide (DCA-) complexes has been developed where f-element salts are dissolved in DCA--based ionic liquids (ILs) directly or formed in situ, forcing coordination of these normally weakly coordinating soft N-donor anions, even in an ambient, non-moisture-excluding environment. A series of lanthanide complexes [C2mim][Ln(DCA)4(H2O)4] (C2mim = 1-ethyl-3-methylimidazolium; Ln = La, Nd, Eu, Tb, Dy, and Yb) and [C2mim]3n[La(OH2)4(µ2-DCA)4]n[La(OH2)2(µ3-DCA)3(µ2-DCA)4]2n(Cl)4n were crystallized under a variety of conditions using this methodology and structurally characterized using single crystal X-ray diffraction. Although not all examples were isostructural, the dominant feature across the series was the presence of [Ln(DCA)4(H2O)4]- anionic nodes with all terminal DCA- ligands accepting hydrogen bonds from the coordinated water molecules forming a 3D metal organic framework. To determine if any structural clues might aid in the further development of the synthetic methodology, the metal-free IL [C1mim][DCA] (C1mim = 1,3-dimethylimidazolium), a room-temperature solid, crystalline analogue of the reaction IL, which is liquid at room temperature, was also prepared and structurally characterized. The ready isolation of these compounds allowed us to begin an investigation of the physical properties such as the luminescence at room and low temperatures for the Eu, Tb, and Dy representatives.

Two-dimensional layered lanthanide diphosphonates: synthesis, structures and sensing properties toward Fe3+ and Cr2O72.

A series of lanthanide diphosphonates, namely Ln(HL)(H2O)2 (Ln = Nd 1, Eu 2, Tb 3 and Er 4), have been synthesized from a semirigid diphosphonate ligand, (5-methyl-1,3-phenylene)bis(methylene)bisphosphonic acid (H4L). These lanthanide diphosphonates have been systematically characterized by using powder and single-crystal X-ray diffraction, elemental analysis, TGA, IR, UV-vis absorption and luminescence techniques. The single-crystal XRD measurements revealed that these compounds all have two-dimensional layered crystal structures. Among these four compounds, 1, 2 and 4 are isostructural and crystallize in the P21/c space group, whereas compound 3 crystallizes in the P21 space group. These compounds display the characteristic emissions of the respective lanthanide ions. The sensing properties of compound 3 were investigated which revealed that it could be used as a luminescent probe for Fe3+ and Cr2O72- with good selectivity and sensitivity.

Probing the molecular toxic mechanism of di-(2-ethylhexyl) phthalate with glutathione transferase Phi8 from Arabidopsis thaliana.

As the most widely used plasticizer, di-(2-ethylhexyl) phthalate (DEHP) has been extensively applied to agriculture. However, its excessive accumulation may cause the active oxygen damage in plants and further lead to the destruction of antioxidant enzymes. As a core component of the glutathione antioxidant enzyme system, glutathione S-transferases (GSTs) have been reported as biomarkers for assessing oxidative damage induced by environmental pollutants, but the underlying toxic molecular mechanism has rarely been exploited. In this article, the interaction mechanism of Arabidopsis thaliana glutathione S-transferase AtGSTF8 and plasticizer DEHP was investigated at the molecular level by multispectral methods. The enzyme activity changes of AtGSTF8 upon binding with DEHP were also evaluated. A single binding site of AtGSTF8 towards DEHP was predicted and the binding force was presumed mainly by Van der Waals' force and hydrogen bonding based on static quenching mechanism. Besides, the deconstructions of the protein skeleton were also deduced based on the multispectral results and the hazardous effects of DEHP on plants growth were further demonstrated. This work will help to clarify the functional mechanism between the plasticizer DEHP and the antioxidant enzyme AtGSTF8 at the molecular level, and providing useful information for further study of the toxic effects of DEHP on plant antioxidant systems.

Glutathione peroxidase 6 from Arabidopsis thaliana as potential biomarker for plants exposure assessment to di-(2-ethylhexyl) phthalate.

As a most abundant plasticizer, Di-(2-ethylhexyl) phthalate (DEHP) has been widely used in agriculture with an associated potential toxicity to many species including plants via the production of the excessive reactive oxygen species (ROS). However, the potential toxic mechanisms of the plasticizer DEHP-induced oxidative damage to plants remain unknown. The antioxidant enzyme glutathione peroxidase has been suggested as biomarkers to reflect over excessive oxidative stress. In this study, the effect of DEHP on AtGPX6 was evaluated by multi-spectroscopic techniques and molecular docking method. The fluorescence intensity of AtGPX6 was reduced by the static quenching mechanism upon the addition of DEHP. The predominant forces in complex formation was mainly impelled by hydrogen bonding and Van der Waals forces based on the negative ΔH and ΔS, which was in accordance with the molecular docking results. In addition, the secondary structural changes resulted from the complex formation were investigated in presence of different amounts of DEHP by the combination of fluorescence, UV-vis absorption and Circular dichroism spectra, which revealed the loosening and unfolding of the framework of AtGPX6 accompanied with the enhancement of the hydrophilicity around the tryptophan residues. The exploration of the interaction mechanism of DEHP with AtGPX6 at molecular level would help to evaluate the toxicity of the plasticizers and forecast the related adverse effects on plants.

Synthesis of Anhydrous Acetates for the Components of Nuclear Fuel Recycling in Dialkylimidazolium Acetate Ionic Liquids.

A series of anhydrous acetate salts with uranium {[C2C1im][UO2(OAc)3] (1), [C2C2im][UO2(OAc)3] (2), and [C4C1im][UO2(OAc)3] (3)}, lanthanides {[C2C2im]2[La(OAc)5] (4) and [C2C1im]2[Nd(OAc)5] (5)}, and strontium {[C2C1im]n[Sr(OAc)3]n (6)} (where C2C1im = 1-ethyl-3-methylimidazolium, C2C2im = 1,3-diethylimidazolium, C4C1im = 1-butyl-3-methylimidazolium, and OAc = acetate) have been prepared and structurally characterized. Both lanthanides and strontium are common components of the nuclear fuel waste, and their separation from uranium is an important but still challenging task. A new synthetic approach with dialkylimidazolium acetate ionic liquids (ILs) as the solvent has been developed for the direct synthesis of homoleptic acetates from the corresponding hydrates and, unexpectedly, hardly soluble f-element oxides. Although the group of characterized compounds shows perfect structural variability, all actinide and lanthanide metal ions form monomeric complex anions where the metal cation coordinates to five ligands including two oxygen atoms in the case of uranium, as is commonly observed for uranyl compounds. Crystallographic analyses revealed that the complex [UO2(OAc)3]- anions possess rather standard D3h symmetry featuring a hexagonal-bipyramidal coordination environment, while the lanthanide anions [Ln(OAc)5]2- are fully asymmetric and the Ln3+ cations are 10-coordinated in the form of a distorted bicapped tetragonal antiprism. This is the first report of lanthanide ions coordinated in this fashion. For Sr2+, 9-fold coordination through oxygen atoms in the form of a strongly distorted tricapped trigonal prism is observed. The crystallization of anhydrous, homoleptic, anionic acetate complexes from such a large variety of different metal salts appears to be due to the properties of dialkylimidazolium acetate ILs themselves, including enhanced basicity from the high concentration of free anions and their greater affinity for hydrogen-bonding solutes relative to metal cations.

Probing the molecular toxic mechanism of lead (II) ions with glutathione peroxidase 6 from Arabidopsis thaliana.

Along with non-biodegradability and accumulation in agricultural soil, lead (II) ions exert considerable harmful effects on plants even at trace amount, especially for the oxidative damages elicited by the lead ions-induced excessive reactive oxygen species (ROS). The glutathione peroxidases were reported to be correspondent with the oxidative stress induced by heavy metals. However, limited data are available about the potential hazardous mechanisms of the lead ions-induced oxidative damage to plants at molecular level. In this study, the harmful impacts of lead ions on Arabidopsis thaliana glutathione peroxidase 6 (AtGPX6) were assessed based on multi-spectroscopic measurements and molecular docking study. The characteristic fluorescence of AtGPX6 was quenched by lead ions with static mechanism at different temperatures. AtGPX6 exhibits a single binding site with lead ions, and then the complex formation was mainly driven by hydrogen bonding interaction and van der Waals forces on account of the negative ΔH and ΔS. The secondary structural changes were observed from the synchronous fluorescence, UV-visible absorption and Circular dichroism spectra, which led to loosen and unfold of the protein framework accompanied by the incremental hydrophobicity around the vicinity of the tryptophan residues. Therefore, this work illustrates the detailed binding mode between lead (II) ions and glutathione peroxidase 6 from Arabidopsis thaliana and the toxic effects on antioxidative defense system induced by lead ions at molecular level.

Highly Luminescent Ionic Liquids Based on Complex Lanthanide Saccharinates.

Four strongly luminescent ionic liquids with complex lanthanide saccharinate anions, [C4mim]3[Eu(Sac)6(H2O)2] (1), {C4mpy}5{[Ln(Sac)6(H2O)2][Ln(Sac)5(H2O)3]}{(H2O)2(CH3CN)2} (Ln = Sm for 2a; Ln = Eu for 2b), and [C4mpy]3[Eu(Sac)6][CH3CN] (3) (C4mim = 1-butyl-3-methylimidazolium; C4mpy = N-butyl-4-methylpyridinium; Sac = saccharinate), have been obtained by reacting the ionic liquids 1-butyl-3-methylimidazolium saccharinate, [C4mim][Sac], and N-butyl-4-methylpyridinium saccharinate, [C4mpy][Sac], with the respective lanthanide saccharinates. Single-crystal X-ray diffraction analyses reveal the respective lanthanide center to be six- or eight-coordinated by five or six saccharinate anions and two or three aqua ligands in the cases of 1 and 2 when lanthanide saccharinated hydrate was employed as the starting material. Coordination of water to the lanthanide can be avoided when using the anhydrous lanthanide saccharinate as shown by the structure of 3. Using a co-solvent, acetonitrile, to facilitate the reaction led to incorporation of solvent molecules into the crystal structure of the final materials (2 and 3). Differential scanning calorimetry analyses reveal that 1 is an ionic liquid whereas 2 and 3 are low-temperature molten salts. The three europium(III)-containing compounds (1, 2b, and 3) all show characteristic intense red emissions of Eu(III) upon excitation into levels of the saccharinate ligands (321 nm) or Eu(III) (393 nm). At room temperature, the decay times of 1 and 2b are all ∼0.5 ms, whereas the decay time of 3 amounts to 3.85 ms due to removal of aqua ligands in the first coordination sphere; accordingly, the quantum efficiencies of 1, 2b, and 3 were determined to be 15.9%, 22.95%, and 57.75%, respectively. The CIE chromaticity coordinates for all Eu compounds are in the red region and approach the NTSC standard CIE values when the temperature is increased. Sm(III)-containing compound 2a shows characteristic Sm(III) emission peaks. As expected, the CIE coordinates of samarium compound 2a fall in the orange-red region.

Molecular mechanism study on the interactions of cadmium (II) ions with Arabidopsis thaliana glutathione transferase Phi8.

Accumulation of cadmium ions may result in adverse effects on plant due to the oxidative stress via destructions of antioxidants and antioxidant enzymes. As the core component of the glutathione antioxidant system, glutathione S-transferases (GSTs) have been reported as biomarkers for evaluating the metal-induced oxidative damage to plants, but the potential toxicity and underlying toxic molecular mechanisms remain unknown. This article investigated the molecular interactions of cadmium ions with Arabidopsis thaliana glutathione S-transferase phi8 (AtGSTF8) by multi-spectroscopic techniques and enzyme activity measurements. The intrinsic fluorescence of AtGSTF8 was quenched statically upon the addition of cadmium ions accompanied with the complex formation and structural and conformational alterations from multiple spectroscopic measurements, resulting in deconstructed protein skeleton and microenvironmental alterations around the Tyr and Trp residues. A single binding site was predicted for AtGSTF8 towards cadmium ions and the van der Walls interactions and hydrogen bonds are the major driving forces of the interaction. In addition, the transferase activity changes of AtGSTF8 upon the addition of cadmium ions have been observed. The implementation of this work helps to clarify the mechanism of oxidative damage and antioxidant enzymes response induced by heavy metal accumulation in plant at molecular level.

Structural Tuning and Sensitization of Uranyl Phosphonates by Incorporation of Countercations into the Framework.

By the introduction of terephthalic acid, tetramethylammonium chloride, and Zn2+ ions, three new uranyl triphosphonates with varying crystal structures, namely, [H3O][(UO2)2(HL)]·H2O (1), [NMe4][(UO2)2(HL)(H2O)]·H2O (2), and [(UO2)3Zn(H2L)2(H2O)2]·3H2O (3), where H6L = benzene-1,3,5-triyltris(methylene) triphosphonic acid, have been successfully synthesized and characterized by means of powder and single crystal XRD, IR, EA, TGA, UV-vis, and luminescence. These three compounds all possess three-dimensional framework structures with hydrium, tetramethylammonium, and Zn2+ as the respective countercations. The uranium(VI) center luminescence of compounds 1 and 2 is completely quenched. However, the incorporation of Zn2+ into the matrix of uranyl phosphonate in the case of compound 3 results in the emerging of the typical intense vibronic emissions of U(VI), demonstrating that zinc phosphonate can behave as sensitizer of uranyl phosphonates. The quenching and sensitization mechanism were also discussed.

Molecular mechanism investigation on the interactions of copper (II) ions with glutathione peroxidase 6 from Arabidopsis thaliana.

Accumulation of copper (II) ions in plant leads to the excessive reactive oxygen species (ROS) which attributes to the depletion of the antioxidants in the cell and destruction to antioxidant enzymes. The antioxidant enzyme glutathione peroxidase has been used as biomarkers to reflect metal-induced oxidative stress. However, the underlying toxic mechanisms of the copper ions(II)-induced oxidative damage to plants remain unknown. In the work, a detailed molecular interaction of copper (II) ions with Arabidopsis thaliana glutathione peroxidase 6 (AtGPX6) in relation with poisonous effects of exposure to heavy metal was investigated by multiple spectroscopic techniques. The intrinsic fluorescence of AtGPX6 was quenched upon the addition of copper (II) ions by the combination of static and dynamic quenching mechanisms accompanied by complex formation and conformational changes. A single binding site was revealed for AtGPX6 towards copper ions. The binding process was hydrophobic effect accompanied by positive entropy change and enthalpy change. The secondary structure of AtGPX6 was changed by the addition of copper ions investigated by synchronous fluorescence spectroscopy, ultraviolet-visible absorption spectroscopy and circular dichroism spectroscopy, resulting in loosened and deconstructed protein skeleton and increased hydrophobicity around the Trp residues. This study helps to illuminate the detailed interactions between copper ions and plant glutathione peroxidase and elucidate the destructive mechanism to antioxidative defense system caused by heavy metal exposure at molecular level.