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Hybrid structures incorporating different organic and inorganic constituents are emerging as a very promising class of materials since they synergistically combine the complementary and diverse properties of the individual components. Hybrid materials based on polyoxometalate clusters (POMs) are particularly interesting due to their versatile catalytic, redox, electronic, and magnetic properties, yet the controlled incorporation of different clusters into a hybrid structure is challenging and has been scarcely reported. Herein we propose a novel and general strategy for combining multiple types of metal-oxo clusters in a single hybrid molecule. Two novel hybrid POM structures (HPOMs) bis-functionalised with dipentaerythritol (R-POM1-R; R = (OCH2)3CCH2OCH2C(CH2OH)) were synthesised as building-blocks for the formation of heterometallic hybrid triads (POM2-R-POM1-R-POM2). Such a modular approach resulted in the formation of four novel heterometallic hybrids combing the Lindqvist {V6}, Anderson-Evans {XMo6} (X = Cr or Al) and trisubstituted Wells-Dawson {P2V3W15} POM structures. Their formation was confirmed by multinuclear Nuclear Magnetic Resonance (NMR), infrared (IR) and UV-Vis spectroscopy, as well as Mass Spectrometry, Diffusion Ordered Spectroscopy (DOSY) and elemental analysis. The thermal stability of the hybrids was also examined by Thermogravimetric Analysis (TGA), which showed that the HPOM triads exhibit higher thermal stability than comparable hybrid structures containing only one type of POM. The one-pot synthesis of these novel compounds was achieved in high yields in aqueous and organic media under simple reflux conditions, without the need of any additives, and could be translated to create other hybrid materials based on a variety of metal-oxo cluster building-blocks. ispartof: CHEMICAL SCIENCE vol:13 issue:10 pages:2891-2899 ispartof: location:England status: published

The performance of MOFs in catalysis is largely derived from structural features, and much work has focused on introducing structural changes such as defects or ligand functionalisation to boost the reactivity of the MOF. However, the effects of different parameters chosen for the synthesis on the catalytic reactivity of the resulting MOF remains poorly understood. Here, we evaluate the role of metal precursor on the reactivity of Zr-based MOF-808 towards hydrolysis of the peptide bond in the glycylglycine model substrate. In addition, the effect of synthesis temperature and duration has been investigated. Surprisingly, the metal precursor was found to have a large influence on the reactivity of the MOF, surpassing the effect of particle size or number of defects. Additionally, we show that by careful selection of the Zr-salt precursor and temperature used in MOF syntheses, equally active MOF catalysts could be obtained after a 20 minute synthesis compared to 24 h synthesis. ispartof: CHEMISTRY-A EUROPEAN JOURNAL vol:27 issue:68 pages:17230-17239 ispartof: location:Germany status: published

The catalytic activity of Zr6-based metal–organic frameworks (Zr-MOFs) toward peptide bond formation is investigated using dipeptide cyclization as a model reaction. Unlike previous catalysts, Zr-MOFs largely tolerate water, reactions are carried out under ambient conditions, the catalyst is recyclable and no additives to activate the COOH group are necessary. The reaction mechanism was assessed by detailed mechanistic and computational studies and features a Lewis acid activation of carboxylate groups by Zr centers toward amine addition in which an alkoxy ligand on adjacent Zr sites assists in lowering the barrier of key proton transfers. The proposed concepts were also used to study the formation of intermolecular peptide bond formation. ispartof: ACS CATALYSIS vol:11 issue:13 pages:7647-7658 status: published

The controlled formation of protein supramolecular assemblies is challenging, but it could provide an important route for the development of hybrid biomaterials. In this work, we demonstrate the fo...

The synthesis and characterization of small monodisperse multimodal iron oxide nanoparticles (IONPs), functionalized with boron-dipyrromethene (BODIPY) derivatives is described. Optical and relaxometric properties of the nanoparticles have been evaluated, suggesting a potential applicability of the particles as bimodal contrast agent for magnetic resonance and optical imaging. The BODIPY funcionalized IONPs showed bright fluorescence and high transverse relaxivity r2 which was in the range 47-49 mM?1 s?1. Two-photon fluorescence microscopy experiments performed at 1100 nm revealed that BODIPY-functionalized IONPs showed significant increase in two-photon fluorescence compared to the bare nanoparticles, suggesting their potential for applications in multimodal imaging. Cell viability studies on rat tumor (AR42J) and human ovarian cancer (SKOV3) cells showed no significant cytotoxicity at any of the tested concentrations, while greater sensitivity was observed for human hepatic stellate (GRX), human umbilical vein (HUVEC) and rat insulinoma (INS1) cells at higher nanoparticle concentrations. https://doi.org/10.1002/nano.202000022 ispartof: Nano select vol:2 issue:2 pages:406-416 status: published

Proteolytic activity of heterogeneous Zr-based nanozymes is a promising technology for the development of selective protein cleavage protocols, which are pivotal in modern proteomics. Here, we repo...

Structural parameters influencing the reactivity of metal-organic frameworks (MOF) are challenging to establish. However, understanding their effect is crucial to further develop their catalytic potential. Here, we uncovered a correlation between reaction kinetics and the morphological structure of MOF-nanozymes using the hydrolysis of a dipeptide under physiological pH as model reaction. Comparison of the activation parameters in the presence of NU-1000 with those observed with MOF-808 revealed the reaction outcome is largely governed by the Zr6 cluster. Additionally, its structural environment completely changes the energy profile of the hydrolysis step, resulting in a higher energy barrier ΔG ‡ for NU-1000 due to a much larger ΔS ‡ term. The reactivity of NU-1000 towards a hen egg white lysozyme protein under physiological pH was also evaluated, and the results pointed to a selective cleavage at only 3 peptide bonds. This showcases the potential of Zr-MOFs to be developed into heterogeneous catalysts for non-enzymatic but selective transformation of biomolecules, which are crucial for many modern applications in biotechnology and proteomics. ispartof: CHEMICAL SCIENCE vol:11 issue:26 pages:6662-6669 ispartof: location:England status: published

ispartof: CRYSTAL GROWTH & DESIGN vol:22 issue:3 status: published

Despite the enormous importance of insoluble proteins in biological processes, their structural investigation remains a challenging task. The development of artificial enzyme-like catalysts would greatly facilitate the elucidation of their structure since currently used enzymes in proteomics largely lose activity in the presence of surfactants, which are necessary to solubilize insoluble proteins. In this study, the hydrolysis of a fully insoluble protein by polyoxometalate complexes as artificial proteases in surfactant solutions is reported for the first time. The hydrolysis of zein as a model protein was investigated in the presence of Zr(IV) and Hf(IV) substituted Keggin-type polyoxometalates (POMs), (Et2 NH2 )10 [M(α-PW11 O39 )2 ] (M = Zr or Hf), and different concentrations of the anionic surfactant sodium dodecyl sulfate (SDS). Selective hydrolysis of the protein upon incubation with the catalyst was observed, and the results indicate that the hydrolytic selectivity and activity of the POM catalysts strongly depends on the concentration of surfactant. The molecular interactions between the POM catalyst and zein in the presence of SDS were explored using a combination of spectroscopic techniques which indicated competitive binding between POM and SDS towards the protein. Furthermore, the formation of micellar superstructures in ternary POM/surfactant/protein solutions has been confirmed by conductivity and Dynamic Light Scattering measurements. ispartof: CHEMISTRY-A EUROPEAN JOURNAL vol:28 issue:8 ispartof: location:Germany status: published

The ability of soluble metal-oxo clusters to specifically interact with protein surfaces makes them attractive as potential inorganic drugs and as artificial enzymes. In particular, metal-substituted polyoxometalates (MS-POMs) are remarkably selective in hydrolyzing a range of different proteins. However, the influence of MS-POMs' redox chemistry on their proteolytic activity remains virtually unexplored. Herein we report a highly site-selective hydrolysis of hemoglobin (Hb), a large tetrameric globular protein, by a Ce(iv)-substituted Keggin polyoxometalate (CeIVK), and evaluate the effect of CeIVK's redox chemistry on its reactivity and selectivity as an artificial protease. At pH 5.0, incubation of Hb with CeIVK resulted in strictly selective protein hydrolysis at six Asp-X bonds, two of which were located in the α-chain (α(Asp75-Leu76) and α(Asp94-Pro95)) and five at the β-chain (β(Asp51-Ala52), β(Asp68-Ser69), β(Asp78-Asp79), β(Asp98-Pro99) and β(Asp128-Phe129)). However, increasing the pH of the reaction mixture to 7.4 decreased the CeIVK hydrolytic reactivity towards Hb, resulting in the cleavage of only one peptide bond (β(Asp128-Phe129)). Combination of UV-Vis, circular dichroism and Trp fluorescence spectroscopy indicated similar interactions between Hb and CeIVK at both pH conditions; however, 31P NMR spectroscopy showed faster reduction of CeIVK into the hydrolytically inactive CeIIIK form in the presence of protein at pH 7.4. In agreement with these results, careful mapping of all hydrolyzed Asp-X bonds on the protein structure revealed that the lower reactivity toward the α-chain was consistent with the presence of more redox-active amino acids (Tyr and His) in this subunit in comparison with the β-chain. This points towards a link between the presence of the redox-active sites on the protein surface and efficiency and selectivity of redox-active MS-POMs as artificial proteases. More importantly, the study provides a way to tune the redox and hydrolytic reactivity of MS-POMs towards proteins through adjustment of reaction parameters like temperature and pH., The redox chemistry of CeIV-polyoxometalates towards proteins is linked to the redox-active residues on protein surface. It can be tuned by adjusting reaction parameters, directly impacting its efficiency and selectivity as an artificial protease.

Materials with enzyme-like activities and proteolytic potential are emerging as a robust and effective alternative to natural enzymes. Herein, a Hf6O8-based NU-1000 metal organic framework (Hf-MOF) is shown to act as a heterogeneous catalyst for the hydrolysis of peptide bonds under mild conditions. In the presence of Hf-MOF, a glycylglycine model dipeptide was hydrolysed with a rate constant of kobs = 8.33 × 10-7 s-1 (half-life (t1/2) of 231 h) at 60 °C and pD 7.4, which is significantly faster than the uncatalyzed reaction. Other Gly-X peptides (X = Ser, Asp, Ile, Ala, and His) were also smoothly hydrolysed under the same conditions with similar rates, except for the faster reactions observed for Gly-His and Gly-Ser. Moreover, the Hf6O8-based NU-1000 MOF also exhibits a high selectivity in the cleavage of a protein substrate, hen egg white lysozyme (HEWL). Our results suggest that embedding Hf6O8 oxo-clusters is an efficient strategy to conserve the hydrolytic activity while smoothing the strong substrate adsorption previously observed for a discrete Hf oxo-cluster that hindered further development of its proteolytic potential. Furthermore, comparison with isostructural Zr-NU-1000 shows that although the Hf variant afforded the same cleavage pattern towards HEWL but slightly slower reaction rates, it exhibited a larger stability window and a better recyclability profile. The results suggest that these differences originate from the intrinsic differences between HfIV and ZrIV centers, and from the lower surface area of Hf-NU-1000 in comparison to Zr-NU-1000. ispartof: NANOSCALE vol:13 issue:28 pages:12298-12305 ispartof: location:England status: published

Single-type, 6-fold symmetrically grooved, and commercially accessible interlocking disks (ILDs) have been used for modeling of sp2 hybridized carbon-based nanoarchitectures and complex polyhedral and reticular material models. In the case of the carbon-based nanoarchitectures, we showcase that the primary ILDs can be directly used for representing individual atoms and bonds. Further on, the spatial connectivity of the primary ILDs can be extended by assembly of symmetrical secondary building units (SBUs). The constructed (deci)meter scale models are robust, light, scalable, and suitable for classroom demonstrations. The ILD technique is also suitable for use in workshops for facile discovery-based learning of nanomolecular structure, showing promise for wider use in the chemistry curriculum.

A simple, safe and inexpensive amide bond formation directly from non-activated carboxylic acids and free amines is presented in this work. Readily available Zr(IV)- and Hf(IV)-substituted polyoxometalates (POM) are shown to be excellent catalysts for the amide bond formation reaction under mild conditions, low catalyst loading, and without the use of water scavengers, dry solvents, additives for facilitating the amine attack, or specialized experimental setups commonly employed to remove water. Detailed mechanistic investigations revealed the key role of polyoxometalate scaffolds which act as inorganic ligands to protect Zr(IV) and Hf(IV) Lewis acidic metals against hydrolysis, and preserve their catalytic activity in amide bond formation reactions. The new catalysts are compatible with a range of functional groups and heterocycles useful for medicinal, agrochemical and material chemists. The robustness of the Lewis acid-polyoxometalate complexes is further supported by the catalyst reuse without loss of activity. This prolific combination of Zr(IV)/Hf(IV) and polyoxometalates inaugurates a powerful class of catalysts for the amide bond formation, which overcomes key limitations of previously established Zr(IV)/Hf(IV) salts and boron based catalysts. ispartof: Acs Catalysis vol:9 issue:11 pages:10245-10252 status: published

Contrast agents for magnetic resonance imaging have historically been based on paramagnetic metal complexes, particularly Gd3+ chelates, which tend to lose their contrast enhancement ability with increasing magnetic field strength. Emerging high-field MRI applications require the development of novel contrast agents that exhibit high relaxation enhancement as a function of magnetic field strength. Paramagnetic ions such as Dy3+ , Tb3+ or Ho3+ incorporated into supramolecular or inorganic nano-architectures represent promising platforms for the development of high field MRI contrast agents. Furthermore, such platforms allow facile inclusion of multiple imaging modalities, therapeutic loading, and targeting vectors. This Minireview examines the application of contrast agents for high-field MRI, which range from single molecules to nanoparticles. Approaches to create multimodal agents by combining high-field MRI contrast properties with another imaging modality are also discussed. ispartof: CHEMISTRY-A EUROPEAN JOURNAL vol:25 issue:61 pages:13838-13847 ispartof: location:Germany status: published

Polyoxometalates have long been studied in a variety of biological applications. Interactions between the highly charged POM molecules and biological molecules frequently occur through hydrogen-bonding and electrostatic interactions. Tellurium-centred Anderson-Evans POMs show exceptional promise as crystallization agents, while acidic and metal-substituted POMs may provide interesting alternatives to enzymes in proteomics applications. While POMs also show interesting results in a number of medicinal applications, for example as anti-amyloid agents for the treatment of Alzheimer's disease and as anti-tumoral agents, their use is often impeded by their toxicity. Many recent studies have therefore focussed on POM-functionalization to reduce toxicity and increase activity by addition of biological targeting molecules. ispartof: CURRENT OPINION IN BIOTECHNOLOGY vol:58 issue:58 pages:92-99 ispartof: location:England status: Published online

Mixed micelles based on amphiphilic gadolinium(III)-DOTA and europium(III)-DTPA complexes were synthesized and evaluated for their paramagnetic and optical properties as potential bimodal contrast agents. Amphiphilic folate molecule for targeting the folate receptor protein, which is commonly expressed on the surface of many human cancer cells, was used in the self-assembly process in order to create nanoaggregates with targeting properties. Both targeted and nontargeted nanoaggregates formed monodisperse micelles having distribution maxima of 10 nm. The micelles show characteristic europium(III) emission with quantum yields of 2% and 1.1% for the nontargeted and targeted micelles, respectively. Fluorescence microscopy using excitation at 405 nm and emission at 575-675 nm was employed to visualize the nanoaggregates in cultured HeLa cells. The uptake of folate-targeted and nontargeted micelles is already visible after 5 h of incubation and was characterized with the europium(III) emission, which is clearly observable in the cytoplasm of the cells. The very fast longitudinal relaxivity r

Estrogen deficiency induces bone loss by increasing bone resorption, in part through upregulation of receptor activator of nuclear factor-κB ligand (RANKL). RANKL is secreted by osteoblasts and osteocytes, but more recently bone marrow (pre)adipocytes have also been shown to express RANKL. Estrogen deficiency increases bone marrow adipose tissue (BMAT). The aim of this study was to determine the effect of ovariectomy (OVX) on RANKL protein expression by bone marrow adipocytes in C3H/HeJ mice. Fourteen-week-old female C3H/HeJ mice ( n = 20) were randomized to sham surgery (Sham) or OVX. After 4 wk animals were euthanized. BMAT volume fraction (BMAT volume/marrow volume) was quantified by polyoxometalate-based contrast-enhanced nano-computed tomography. The percentage of RANKL-positive bone marrow adipocytes (RANKL-positive bone marrow adipocytes/total adipocytes) and the percentage of RANKL-positive osteoblasts covering the bone surface (bone surface covered in RANKL-positive osteoblasts/total bone surface) were quantified in the distal metaphysis of immunohistochemically stained sections of the left femur. The effects of OVX were analyzed by Student’s t test or Mann–Whitney U test. RANKL was detected in osteoblasts, osteocytes, and bone marrow adipocytes. OVX significantly increased mean percentage of RANKL-positive bone marrow adipocytes [mean (SD): Sham 42 (18)%; OVX 64 (12)%; P = 0.029] as well as BMAT volume/marrow volume [median (interquartile range): Sham 1.4 (4.9)%; OVX 7.2 (7.3)%; P = 0.008] compared with Sham. We show that OVX increased both the percentage of RANKL-positive bone marrow adipocytes and the total BMAT volume fraction in C3H/HeJ mice. Therefore, RANKL produced by bone marrow adipocytes could be an important contributor to OVX-induced bone loss in C3H/HeJ mice.

The discovery of nanozymes for selective fragmentation of proteins would boost the emerging areas of modern proteomics, however, the development of efficient and reusable artificial catalysts for peptide bond hydrolysis is challenging. Here we report the detailed catalytic properties of a microporous zirconium carboxylate metal-organic framework, MIP-201, in promoting peptide bond hydrolysis in a simple dipeptide, as well as in horse-heart myoglobin (Mb) protein that consists of 153 amino acids. We demonstrate that MIP-201 features an excellent catalytic activity and selectivity, a good tolerance toward reaction conditions covering a wide range of different pH values, and importantly, an exceptional recycling ability associated with easy regeneration process. Taking into account the excellent catalytic performance of MIP-201 and its other advantages such as 6-connected Zr6 cluster active sites, the green, scalable and cost-effective synthesis, and an outstanding chemical and architectural stability, our finding suggests that MIP-201 may be a promising and practical alternative to the current commercially available catalysts for peptide bond hydrolysis.

The catalytic activity of metal–organic frameworks (MOFs) toward peptides and proteins provides an attractive route for the development of nanozymes for applications in biotechnology and proteomics...

Understanding the stability and speciation of metal-oxo clusters in solution is essential for many of their applications in different areas. In particular, hybrid organic-inorganic polyoxometalates (HPOMs) have been attracting increasing attention as they combine the complementary properties of organic ligands and metal-oxygen nanoclusters. Nevertheless, the speciation and solution behavior of HPOMs have been scarcely investigated. Hence, in this work, a series of HPOMs based on the archetypical Anderson-Evans structure, δ-[MnMo6O18{(OCH2)3C-R}2]3-, with different functional groups (R = -NH2, -CH3, -NHCOCH2Cl, -N═CH(2-C5H4N) {pyridine; -Pyr}, and -NHCOC9H15N2OS {biotin; -Biot}) and countercations (tetrabutylammonium {TBA}, Li, Na, and K) were synthesized, and their solution behavior was studied in detail. In aqueous solutions, decomposition of HPOMs into the free organic ligand, [MoO4]2-, and free Mn3+ was observed over time and was shown to be highly dependent on the pH, temperature, and nature of the ligand functional group but largely independent of ionic strength or the nature of the countercation. Furthermore, hydrolysis of the amide and imine bonds often present in postfunctionalized HPOMs was also observed. Hence, HPOMs were shown to exhibit highly dynamic behavior in solution, which needs to be carefully considered when designing HPOMs, particularly for biological applications. ispartof: INORGANIC CHEMISTRY vol:60 issue:14 pages:10215-10226 ispartof: location:United States status: published

Materials which synergistically combine Lewis and Bronsted acid properties hold potential for applications as efficient heterogeneous catalysts for preparation of biofuels and biolubricants. Herein we report new heterogeneous catalysts based on grafting the intact Lewis metal (ZrIV) substituted Keggin polyoxometalate (POM) on mesoporous silica support. A new approach for immobilization of the POM in MCM-41 silica was developed by co-condensation of Si source (tetraethyl orthosilicate, TEOS) with POM salt in the presence of a template molecule as an alternative to the commonly used acidic POM form and impregnation procedure for catalyst preparation. The proposed synthesis method in combination with extraction of the template proceeded with preservation of the intact POM structure and resulted in hybrid catalysts with in situ generated Bronsted acid sites in addition to the Lewis acidity provided by the metal centers. Textural properties of the catalysts were characterized by X-ray diffraction, N2 physisorption and transmission electron microscopy (TEM). Insight into POM stability and structural transformations during synthesis, template removal and impregnation was provided by solid state 31P and 29Si NMR spectroscopy. Catalytic activity was studied in esterification reactions of levulinic acid with ethanol or octanol to value-added esters. The directly synthesized POM-functionalized hybrid catalysts exceeded the post synthesis impregnated ones, demonstrating significantly higher catalytic activity, recyclability and resistance against leaching. The proposed approach for immobilization of Lewis metal POMs in MCM-41 silica framework with in situ generation of the active Bronsted acid sites opens prospects for the development of efficient hybrid catalysts for esterification reaction, which overcomes the main limitations of common POM based catalysts such as low stability of the acid sites during the synthesis and the catalytic reaction, low surface area, agglomeration of the catalytically active phase and low stability of the Lewis metal center in presence of water.

The controlled formation of protein supramolecular assemblies is challenging but it could provide an important route for the development of hybrid biomaterials. In this work, we demonstrate formation of well-defined complexes formed between the 8-fold symmetrical designer protein Tako8 and soluble metal-oxo clusters from the family of Anderson-Evans, Keggin and ZrIV- substituted Wells-Dawson polyoxometalates. A combination of x-ray crystallography and solution studies showed that metal-oxo clusters are able to serve as linker nodes for the bottom-up creation of protein based supramolecular assemblies. Our findings indicate that clusters with larger size and negative charge are capable of modulating the crystal packing of the protein, highlighting the need for a size and shape complementarity with the protein node for optimal alteration of the crystalline self-assembly.

Efficient and selective hydrolysis of inert peptide bonds is of paramount importance. MOF-808, a metal-organic framework based on Zr6 nodes, can hydrolyze peptide bonds efficiently under biologically relevant conditions. However, the details of the catalyst structure and of the underlying catalytic reaction mechanism are challenging to establish. By means of DFT calculations we first investigate the speciation of the Zr6 nodes and identify the nature of ligands that bind to the Zr6O8H4-x core in aqueous conditions. The core is predicted to strongly prefer a Zr6O8H4 protonation state and to be predominantly decorated by bridging formate ligands, giving Zr6(μ3-O)4(μ3-OH)4(BTC)2(HCOO)6 and Zr6(μ3-O)4(μ3-OH)4(BTC)2(HCOO)5(OH)(H2O) as the most favorable structures at physiological pH. The GlyGly peptide can bind MOF in several different ways, with the preferred structure involving coordination through the terminal carboxylate analogously to the binding mode of formate ligand. The pre-reactive binding mode in which the amide carbonyl oxygen coordinates the metal core lies 7 kcal higher in free energy. The preferred reaction pathway is predicted to have two close-lying transition states, either of which could be the rate-determining step: nucleophilic attack on the amide carbon atom and C-N bond breaking, with calculated relative free energies of 31 and 32 kcal mol-1, respectively. Replacement of formate by water and hydroxide at the Zr6 node is predicted to be possible, but does not appear to play a role in the hydrolysis mechanism. ispartof: PHYSICAL CHEMISTRY CHEMICAL PHYSICS vol:22 issue:43 pages:25136-25145 ispartof: location:England status: published

The development of artificial proteases is challenging, but important for many applications in modern proteomics and biotechnology. The hydrolysis of hydrophobic or unstructured proteins is particularly difficult due to their poor solubility, which often requires the presence of surfactants. Herein, it is shown that a zirconium(IV)-substituted Keggin polyoxometalate (POM), (Et2 NH2 )10 [Zr(α-PW11 O39 )2 ] (1), is able to selectively hydrolyze β-casein, which is an intrinsically unstructured protein at pH 7.4 and 60 °C. Four surfactants (sodium dodecyl sulfate (SDS), N-dodecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (ZW3-12), 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), and polyethylene glycol tert-octylphenyl ether (TX-100)), which differ in the nature of their polar groups, were investigated for their role in influencing the selectivity and efficiency of protein hydrolysis. Under experimental conditions, β-casein forms micellar structures in which the hydrophilic part of the protein is water accessible and able to interact with 1. Identical fragmentation patterns of β-casein in the presence of 1 were observed through SDS poly(acrylamide) gel electrophoresis both in the presence and absence of surfactants, but the rate of hydrolysis varied, depending on the nature of surfactant. Whereas TX-100 surfactant, which has a neutral polar head, caused only a slight decrease in the hydrolysis rate, stronger inhibition was observed in the presence surfactants with charges in their polar heads (CHAPS, ZW3-12, SDS). These results were consistent with those of tryptophan fluorescencequenching studies, which showed that the binding between β-casein and 1 decreased with increasing repulsion between the POM and the polar heads of the surfactants. In all cases, the micellar structure of β-casein was not significantly affected by the presence of POM or surfactants, as indicated by circular dichroism spectroscopy. ispartof: CHEMISTRY-A EUROPEAN JOURNAL vol:26 issue:49 pages:11170-11179 ispartof: location:Germany status: published

The reactivity of a range of Keggin and Wells-Dawson type heteropolyacids (HPAs): H3 PW12 O40 H4 SiW12 O40 , H3 PMo12 O40 , K6 P2 W18 O62 , and NaH2 W12 O4 , towards the heavily glycosylated α-1-acid glycoprotein (AGP) is reported. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) show that after incubation of the protein with HPAs at 80 °C and pH 2.8 complete hydrolysis of terminal glycosidic bond has been achieved, resulting in the removal of sialic acids with no observed destruction of the protein core or the residual glycan chains. The 1 H NMR spectroscopy confirmed that the released sialic acids preserve intact structure upon their excision from the protein, which makes the reported method suitable for the analysis of sialic acid modifications which play an important role in numerous biological processes. The presence of other sugars was not detected by 1 H NMR and HPAEC-PAD, suggesting that HPAs hydrolyze only the terminal glycosidic bond in the glycoprotein, resulting in the selective release of sialic acid from AGP. The kinetic results have shown that under equal temperature and pH conditions, the hydrolysis of the terminal glucosidic bond occurred faster in the presence of HPAs compared to conventional mineral acids. The observed rate constants were in the range 6,7×10-2 -11,9×10-2 min-1 and the complete and selective excision of sialic acids could be achieved within 60 min of incubation. The Trp fluorescence and CD spectroscopy show that non-covalent interaction between HPA and protein takes place in solution which could lead to stabilization of the sialosyl cation that is formed during the glycosidic bond hydrolysis by anionic HPA cluster.

Redox reactions between polyoxometalates (POMs) and biologically relevant molecules have been virtually unexplored but are important, considering the growing interest in the biological applications of POMs. In this work we give a detailed account on the redox behavior of CeIV-substituted polyoxometalates (CeIV-POMs) toward a range of amino acids and peptides. CeIV-POMs have been shown to act as artificial proteases that promote the selective hydrolysis of peptide bonds. In presence of a protein, a concomitant reduction of CeIV to CeIII ion is frequently observed, leading us to examine the origins of this redox reaction by first using amino acid building blocks as simple models. Among all of the examined amino acids, cysteine (Cys) showed the highest activity in reducing CeIV-POMs to CeIII-POMs, followed by the aromatic amino acids tryptophan (Trp), tyrosine (Tyr), histidine (His), and phenylalanine (Phe). While the redox reaction with Cys afforded the well-defined product cystine, no oxidation products were detected for the Trp, His, Tyr, and Phe amino acids after their reaction with CeIV-POMs, suggesting a radical pathway in which the solvent likely regenerates the amino acid. In general, the rate of redox reactions increased upon increasing the pD, temperature, and ionic strength of the reaction. Moreover, the redox reaction is highly sensitive to the type of polyoxometalate scaffold, as complexation of CeIV to a Keggin (K) or Wells-Dawson (WD) polyoxotungstate anion resulted in a large difference in the rate of redox reaction for both Cys and aromatic amino acids. The reduction of CeIVK was at least 1 order of magnitude faster in comparison to CeIVWD, in accordance with the higher redox potential of CeIVK in comparison to CeIVWD. The reaction of CeIVPOMs with a range of peptides containing redox-active amino acids revealed that the redox reaction is influenced by their coordination mode with CeIV ion, but in all examined peptides the redox reaction is favored in comparison to the hydrolytic cleavage of the peptide bond. ispartof: INORGANIC CHEMISTRY vol:59 issue:15 pages:10569-10577 ispartof: location:United States status: published

The selective hydrolysis of proteins by non‐enzymatic catalysis is difficult to achieve, yet it is crucial for applications in biotechnology and proteomics. Herein, we report that discrete hafnium metal‐oxo cluster [Hf18O10(OH)26(SO4)13⋅(H2O)33] (Hf18), which is centred by the same hexamer motif found in many MOFs, acts as a heterogeneous catalyst for the efficient hydrolysis of horse heart myoglobin (HHM) in low buffer concentrations. Among 154 amino acids present in the sequence of HHM, strictly selective cleavage at only 6 solvent accessible aspartate residues was observed. Mechanistic experiments suggest that the hydrolytic activity is likely derived from the actuation of HfIV Lewis acidic sites and the Brønsted acidic surface of Hf18. X‐ray scattering and ESI‐MS revealed that Hf18 is completely insoluble in these conditions, confirming the HHM hydrolysis is caused by a heterogeneous reaction of the solid Hf18 cluster, and not from smaller, soluble Hf species that could leach into solution. This is the peer-reviewed version of the following article: Moons, J.; de Azambuja, F.; Mihailović, J.; Kozma, K.; Smiljanić, K.; Amiri, M.; Ćirković-Veličković, T.; Nyman, M.; Parac-Vogt, T. Discrete Hf18 Metal‐oxo Cluster as a Heterogeneous Nanozyme for Site‐Specific Proteolysis. Angewandte Chemie (International Edition) 2020, 59 (17), 1–9. [https://doi.org/10.1002/anie.202001036]. Supplementary material: [https://cherry.chem.bg.ac.rs/handle/123456789/4851]

The development of modular platforms that can undergo postfunctionalization reactions permits coupling of inorganic clusters with different organic functionalities, thereby expanding the range of key physicochemical properties that are relevant for applications in different areas of science. In this work, a novel hybrid Wells-Dawson polyoxometalate (POM) platform was developed and successfully used for postfunctionalization via a nucleophilic substitution reaction. Two new halogen-functionalized bis-organosilyl Wells-Dawson POMs TBA6[α2-P2W17O61{O(SiC3H6-X)2}] (X = Cl or I) were synthesized, and their coupling with amine substrates was explored in a one-step postfunctionalization reaction. The iodide form of the POM has proven to be much more reactive, and its reaction with a range of primary and secondary amines resulted in a series of new bis-substituted Wells-Dawson POMs with the general formula TBA6[α2-P2W17O61{O(SiC3H6-NR1R2)2}]. Coupling of 18 amines with R1 and R2 groups, which exhibited a wide variety in terms of both chemical nature and bulkiness, was achieved under mild conditions via a catalyst-free approach. Using Na2CO3 as a base in acetonitrile solutions at 55 °C resulted in hybrid products that were obtained in high purity and good yields, after a simple isolation and purification procedure. ispartof: INORGANIC CHEMISTRY vol:59 issue:14 pages:10146-10152 ispartof: location:United States status: published

Acetylcholinesterase (AChE) inhibitors are important in the treatment of neurodegenerative diseases. Two inhibitors, 12-tungstosilicic acid (WSiA) and 12-tungstophosphoric acid (WPA), which have polyoxometalate (POM) type structure, have been shown to inhibit AChE activity in nM concentration. Circular dichroism and tryptophan fluorescence spectroscopy demonstrated that the AChE inhibition was not accompanied by significant changes in the secondary structure of the enzyme. The molecular docking approach has revealed a new allosteric binding site, termed β-allosteric site (β-AS), which is considered responsible for the inhibition of AChE by POMs. To the best of our knowledge, this is the first study reporting a new allosteric site that is considered responsible for AChE inhibition by voluminous and negatively charged molecules such as POMs. The selected POMs were further subjected to genotoxicity testing using human peripheral blood cells as a model system. It was shown that WSiA and WPA induced a mild cytostatic but not genotoxic effects in human lymphocytes, which indicates their potential to be used as medicinal drugs. The identification of non-toxic compounds capable of binding to an allosteric site that so far has not been considered responsible for enzyme inhibition could be fundamental for the development of new drug design strategies and the discovery of more efficient AChE modulators. ispartof: EUROPEAN JOURNAL OF PHARMACEUTICAL SCIENCES vol:151 ispartof: location:Netherlands status: published

MOF-808, a Zr(IV)-based metal-organic framework, has been proven to be a very effective heterogeneous catalyst for the hydrolysis of the peptide bond in a wide range of peptides and in hen egg white lysozyme protein. The kinetic experiments with a series of Gly-X dipeptides with varying nature of amino acid side chain have shown that MOF-808 exhibits selectivity depending on the size and chemical nature of the X side chain. Dipeptides with smaller or hydrophilic residues were hydrolyzed faster than those with bulky and hydrophobic residues that lack electron rich functionalities which could engage in favorable intermolecular interactions with the btc linkers. Detailed kinetic studies performed by 1H NMR spectroscopy revealed that the rate of glycylglycine (Gly-Gly) hydrolysis at pD 7.4 and 60 °C was 2.69 × 10-4 s-1 ( t1/2 = 0.72 h), which is more than 4 orders of magnitude faster compared to the uncatalyzed reaction. Importantly, MOF-808 can be recycled several times without significantly compromising the catalytic activity. A detailed quantum-chemical study combined with experimental data allowed to unravel the role of the {Zr6O8} core of MOF-808 in accelerating Gly-Gly hydrolysis. A mechanism for the hydrolysis of Gly-Gly by MOF-808 is proposed in which Gly-Gly binds to two Zr(IV) centers of the {Zr6O8} core via the oxygen atom of the amide group and the N-terminus. The activity of MOF-808 was also demonstrated toward the hydrolysis of hen egg white lysozyme, a protein consisting of 129 amino acids. Selective fragmentation of the protein was observed with 55% yield after 25 h under physiological pH.

Biological tissues have a complex and heterogeneous 3D structure, which is only partially revealed by standard histomorphometry in 2D. We here present a novel chemical compound for contrast-enhanced microfocus computed tomography (CE-CT), a Hafnium-based Wells-Dawson polyoxometalate (Hf-POM), which allows simultaneous 3D visualization of mineralized and non-mineralized skeletal tissues, such as mineralized bone and bone marrow vasculature and adipocytes . We validated the novel contrast agent, which has a neutral pH in solution, by detailed comparison with (immuno)histology on murine long bones as blueprint , and showed that Hf-POM-based CE-CT can be used for virtual 3D histology. Furthermore, we quantified the 3D structure of the different skeletal tissues, as well as their spatial relation to each other, during aging and diet-induced obesity. We discovered, based on a single CE-CT dataset per sample, clear differences between the groups in bone structure, vascular network organization, characteristics of the adipose tissue and proximity of the different tissues to each other. These findings highlight the complementarity and added value of Hf-POM-based CE-CT compared to standard histomorphometry. As this novel technology provides a detailed 3D simultaneous representation of the structural organization of mineralized bone and bone marrow vasculature and adipose tissue, it will enable to improve insight in the interactions between these three tissues in several bone pathologies and to evaluate the in vivo performance of biomaterials for skeletal regeneration.

Lanthanides, holmium(iii), dysprosium(iii), and terbium(iii), were coordinated to an amphiphilic DOTA bis-coumarin derivative and then further assembled with an amphiphilic europium(iii) DTPA bis-coumarin derivative into mono-disperse micelles. The self-assembled micelles were characterized and assessed for their potential as bimodal contrast agents for high field magnetic resonance and optical imaging applications. All micelles showed a high transverse relaxation (r2) of 46, 34, and 30 s-1 mM-1 at 500 MHz and 37 °C for Dy(iii), Ho(iii) and Tb(iii), respectively, which is a result of the high magnetic moment of these lanthanides and the long rotational correlation time of the micelles. The quantum yield in aqueous solution ranged from 1.8% for Tb/Eu to 1.4% for Dy/Eu and 1.0% for the Ho/Eu micelles. Multi-photon excited emission spectroscopy has shown that due to the two-photon absorption of the coumarin chromophore the characteristic Eu(iii) emission could be observed upon excitation at 800 nm, demonstrating the usefulness of the system for in vivo fluorescence imaging applications. To the best of our knowledge, this is the first example reporting the potential of a holmium(iii) chelate as a negative MRI contrast agent. ispartof: DALTON TRANSACTIONS vol:47 issue:31 pages:10646-10653 ispartof: location:England status: published

A novel, reversible redox-active MRI probe, GdNR1, has been developed for the study of redox changes associated with diseased states. This system exhibits switching in relaxivity upon reduction and oxidation of the appended nicotinimidium. Relaxivity studies and cyclic voltammetry confirmed the impressive reversibility of this system, at a biologically-relevant reduction potential. A 2.5-fold increase in relaxivity was observed upon reduction of the complex, which corresponds to a change in the number of inner-sphere water molecules, as confirmed by luminescence lifetimes of the Eu(iii) analogue and NMRD studies. This is the first example of a redox-responsive MRI probe utilising the biologically-inspired nicotinimidium redox switch. In the future this strategy could enable the non-invasive identification of hypoxic tissue and related cardiovascular disease. ispartof: CHEMICAL COMMUNICATIONS vol:54 issue:92 pages:12986-12989 ispartof: location:England status: published

The reactivity of polyoxovanadates towards adenosine-5′-triphosphate (ATP) hydrolysis at pH 2, 4, 6 and 7 is reported. Detailed kinetic investigation of ATP hydrolysis in the presence of polyoxovanadates was performed through multinuclear nuclear magnetic resonance (NMR) spectroscopy. In general, rate acceleration of up to five orders of magnitude was observed in the presence of vanadates compared to spontaneous ATP hydrolysis, with the greatest acceleration observed for reactions carried out at pH 2. Interestingly, the effectiveness of vanadates in promoting ATP hydrolysis decreased as the pH of the reaction solution increased, nevertheless, at pH = 7, the rate increase of one order of magnitude in comparison to blank reactions was still observed. Interactions between vanadate species in solution and ATP were investigated by means of 31P and 51V NMR spectroscopy, and this pointed towards the preferential interaction of vanadium with the phosphate groups rather than other regions of the ATP molecule.

Group (IV) metal oxo clusters are a diverse class of compounds with rapidly growing applications in catalysis, materials chemistry, electronics and optics. They possess unique structural stability, reactivity and electronic properties; however, their potential remains underexplored due to the gaps in fundamental understanding about their structure, intrinsic features, and nuances in their synthesis that overall would allow for great tunability of their properties. In this context, here we review the chemistry of discrete zirconium, hafnium, and titanium metal oxo clusters reported in the literature in order to provide a critical overview of the structural features, properties and reactivity disclosed to date. While a comprehensive summary of Zr and Hf clusters is presented, only the most recent Ti oxo clusters are discussed in addition to some key compounds that have been revised elsewhere. Envisioning the growing relevance group (IV) metal oxo clusters for the synthesis of nanostructure materials and in catalysis, we also summarize key principles of their surface chemistry, and the photoactivity of Ti oxo clusters, given the paramount importance of these properties for the development of future applications.

This study represents the first example of protein hydrolysis at pH = 7.4 and 60 °C by a metal-substituted polyoxometalate (POM) in the presence of a zwitterionic surfactant. Edman degradation results show that in the presence of 0.5% w/v 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) detergent, a Zr(IV)-substituted Wells-Dawson-type POM, K15H[Zr(α2-P2W17O61)2]·25H2O (Zr1-WD2), selectively hydrolyzes human serum albumin exclusively at peptide bonds involving Asp or Glu residues, which contain carboxyl groups in their side chains. The selectivity and extent of protein cleavage are tuned by the CHAPS surfactant by an unfolding mechanism that provides POM access to the hydrolyzed peptide bonds. ispartof: ACS Omega vol:2 issue:5 pages:2026-2033 ispartof: location:United States status: published

Na/K-ATPase is emerging as an important target for a variety of anticancer metal-based drugs. The interactions of Na/K-ATPase (in its E1 state) with three representative and structurally related cytotoxic gold(III) complexes, i.e. [Au(bipy)(OH)(2)][PF6], bipy = 2,2'-bipyridine; [Au(py(dmb)-H)(CH3COO)(2)], py(dmb)-H = deprotonated 6-(1,1-dimethylbenzyl)-pyridine and [Au(bipy(dmb)-H)(OH)][PF6], bipy(c)-H = deprotonated 6-(1,1-dimethylbenzyl)-2,20-bipyridine, are investigated here in depth using a variety of spectroscopic methods, in combination with docking studies. Detailed information is gained on the conformational and structural changes experienced by the enzyme upon binding of these gold(III) complexes. The quenching constants of intrinsic enzyme fluorescence, the fraction of Trp residues accessible to gold(III) complexes and the reaction stoichiometries were determined in various cases. Specific hypotheses are made concerning the binding mode of these gold(III) complexes to the enzyme and the likely binding sites. Differences in their binding behaviour toward Na/K-ATPase are explained on the ground of their distinctive structural features. The present results offer further support to the view that Na/K-ATPase may be a relevant biomolecular target for cytotoxic gold(III) compounds of medicinal interest and may thus be involved in their overall mode of action.

A mild post-functionalization method, based on a copper(i) catalyzed azide-alkyne cycloaddition, was developed for the fragile [P2W15V3]9- (PWV). PWV was functionalized with an azide and used as a platform to attach various alkyne functionalized compounds.

A highly negatively charged binuclear ZrIV-substituted Keggin polyoxometalate [{α-PW11O39Zr(μ-OH)(H2O)}2]8- (ZrK 2 : 2) has been shown to promote the hydrolytic cleavage of phosphoester bonds in the supercoiled plasmid pUC19 DNA under physiological pH and temperature, giving relaxed and linear forms of pUC19 as hydrolysis products. The interaction between ZrK 2 : 2 and DNA was experimentally proven by circular dichroism (CD) spectroscopy and 31P diffusion ordered NMR spectroscopy. crosscheck: This document is CrossCheck deposited related_data: Supplementary Information identifier: T. N. Parac-Vogt (ORCID) copyright_licence: The Royal Society of Chemistry has an exclusive publication licence for this journal history: Received 24 October 2016; Accepted 9 December 2016; Accepted Manuscript published 9 December 2016; Advance Article published 16 December 2016; Version of Record published 3 January 2017 ispartof: Chemical Communications vol:53 issue:3 pages:617-620 ispartof: location:England status: published

To advance clinical translation of regenerative medicine, there is, amongst others, still need for better insights in tissue development and disease. For this purpose, more precise imaging of the 3D microstructure and spatial interrelationships of the different tissues within organs is crucial. Despite being destructive towards the sample, conventional histology still is the gold standard for structural analysis of biological tissues. It is, however, limited by 2D sections of a 3D object, prohibiting full 3D structural analysis. MicroCT has proven to provide full 3D structural information of mineralized tissues and dense biomaterials. However, the intrinsic low X-ray absorption of soft tissues requires contrast-enhancing staining agents (CESAs). In a previous study, we showed that hafnium-substituted Wells-Dawson polyoxometalate (Hf-WD POM) allows simultaneous contrast-enhanced microCT (CE-CT) visualization of bone and its marrow vascularization and adiposity. In this study, other POM species have been examined for their potential as soft tissue CESAs. Four Wells-Dawson POMs, differing in structure and overall charge, were used to stain murine long bones and kidneys. Their staining potential and diffusion rate were compared to those of Hf-WD POM and phosphotungstic acid (PTA), a frequently used but destructive CESA. Monolacunary Wells-Dawson POM (Mono-WD POM) showed similar soft tissue enhancement as Hf-WD POM and PTA. Moreover, Mono-WD POM is less destructive, shows a better diffusion than PTA, and its synthesis requires less time and cost than Hf-WD POM. Finally, the solubility of Mono-WD POM was improved by addition of lithium chloride (LiCl) to the staining solution, enhancing further the soft tissue contrast. Statement of Significance To advance clinical translation of regenerative medicine, there is, amongst others, still need for better insights in tissue development and disease. For this purpose, more precise imaging of the 3D microstructure and spatial interrelationships of the different tissues within organs is crucial. Current standard structural analysis techniques (e.g. 2D histomorphometry), however, do not allow full 3D assessment. Contrast-enhanced X-ray computed tomography has emerged as a powerful 3D structural characterization tool of soft biological tissues. In this study, from a library of Wells Dawson polyoxometalates (WD POMs), we identified monolacunary WD POM together with lithium chloride, dissolved in phosphate buffered saline, as the most suitable contrast-enhancing staining agent solution for different biological tissues without tissue shrinkage.

Creating efficient and residue-directed artificial proteases is a challenging task due to the extreme inertness of the peptide bond, combined with the difficulty of achieving specific interactions between the catalysts and the protein side chains. Herein we report strictly site-selective hydrolysis of a multi-subunit globular protein, hemoglobin (Hb) from bovine blood, by a range of ZrIV -substituted polyoxometalates (Zr-POMs) in mildly acidic and physiological pH solutions. Among 570 peptide bonds in Hb, selective cleavage was observed at only eleven sites, each occurring at Asp-X peptide bonds located in the positive patches on the protein surface. The molecular origins of the observed Asp-X selectivity were rationalized by means of molecular docking, DFT-based binding, and mechanistic studies on model peptides. The proposed mechanism of hydrolysis involves coordination of the amide oxygen to ZrIV followed by a direct nucleophilic attack of the side chain carboxylate group on the C-terminal amide carbon atom with formation of a cyclic anhydride, which is further hydrolyzed to give the reaction products. The activation energy for the cleavage of the structurally related Glu-X sequence compared to Asp-X was calculated to be higher by 1.4 kcal mol-1 , which corresponds to a difference of about one order of magnitude in the rates of hydrolysis. The higher activation energy is attributed to the higher strain present in the six-membered ring of glutaric anhydride (Glu-X), as compared to the five-membered ring of the succinic anhydride (Asp-X) intermediate. Similarly, the cleavage at X-Asp and X-Glu bonds are predicted to be kinetically less likely as the corresponding activation energies were 6 kcal mol-1 higher, explaining the experimentally observed selectivity. The synergy between the negatively charged polyoxometalate cluster, which binds at positive patches on protein surfaces, and selective activation of Asp-X peptide bonds located in these regions by ZrIV ions, results in a novel class of artificial proteases with aspartate-directed reactivity, which is very rare among naturally occurring proteases. ispartof: CHEMISTRY-A EUROPEAN JOURNAL vol:25 issue:63 pages:14370-14381 ispartof: location:Germany status: published

Hydrolysis of Transferin (Tf), a glycoprotein consisting of 679 amino acids and three glycan chains, has been examined in the presence of Ce(IV)-substituted Keggin polyoxometalate [CeIV(α-PW11O39)2]10− (Ce-K POM). Incubation at pH = 7.4 and at 37 °C resulted in selective fragmentation of the Tf after 24 h, which was followed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE). After 5 days of incubation nearly 90% of Tf was hydrolyzed, giving 9 fragments with molecular weight (Mw) in the range between 13.7 and 69.5 kDa. Under the same conditions, the sample of Tf which did not contain the Ce-K POM, showed only 7% of background hydrolysis. The interactions between Ce-K and Tf were studied by tryptophan fluorescence, circular dichroism (CD) and 31P Nuclear magnetic resonance (NMR) spectroscopy. The association constant (Ka) for the interaction was calculated to be 3.2 × 104 M−1 from tryptophan quenching studies. CD spectroscopy revealed that binding of Ce-K to Tf resulted in significant secondary structure changes, mainly affecting the alpha-helical content of the protein. 31P NMR spectroscopy showed that in the presence of the Tf partial reduction of Ce(IV) to Ce(III) occurred, which did not affect the overall structure of the Keggin POM. ispartof: Polyhedron vol:170 pages:570-575 status: published

© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim Four Wells–Dawson type metal-substituted polyoxometalates (MSPs), 1:2 ZrIV-Wells–Dawson [ZrIV(α2-P2W17O61)]8– (1), 1:1 CoII-Wells–Dawson [CoII(α2-P2W17O61)]10– (2), 1:1 NiII-Wells–Dawson [NiII(α2-P2W17O61)]10– (3) and 1:1 CuII-Wells–Dawson [CuII(α2-P2W17O61)]10– (4), which differ in the nature of the imbedded metal ion, were examined in co-crystallization experiments with a protein Hen Egg White Lysozyme (HEWL). Single crystal X-ray structures of four noncovalent complexes between POMs and HEWL have been determined, and the influence of the type of substituted metal on the mode of POM binding to a protein was investigated. All crystal structures exhibited a high degree of similarity, suggesting that the interaction is largely independent on the nature of substituted metal within the same polyoxometalate (POM) archetype. The main driving force for the formation of the noncovalent complex is electrostatic attraction between POM and HEWL surface regions. Stabilization is further provided by direct and water mediated hydrogen bonding between terminal oxygen atoms of the POM framework and flexible HEWL residues. ispartof: EUROPEAN JOURNAL OF INORGANIC CHEMISTRY vol:2019 issue:3-4 pages:506-511 status: published