Synthesis of Terpyridine-Terminated Amphiphilic Block Copolymers and Their Self-Assembly into Metallo-Polymer Nanovesicles.

Polystyrene-b-polyethylene glycol (PS-b-PEG) amphiphilic block copolymers featuring a terminal tridentate N,N,N-ligand (terpyridine) were synthesized for the first time through an efficient route. In this approach, telechelic chain-end modified polystyrenes were produced via reversible addition-fragmentation chain-transfer (RAFT) polymerization by using terpyridine trithiocarbonate as the chain-transfer agent, after which the hydrophilic polyethylene glycol (PEG) block was incorporated into the hydrophobic polystyrene (PS) block in high yields via a thiol-ene process. Following metal-coordination with Mn2+, Fe2+, Ni2+, and Zn2+, the resulting metallo-polymers were self-assembled into spherical, vesicular nanostructures, as characterized by dynamic light scattering and transmission electron microscopy (TEM) imaging.

Effects of a strong π-accepting ancillary ligand on the water oxidation activity of weakly coupled binuclear ruthenium catalysts.

Significant differences were found in the proton-coupled redox chemistry and catalytic behavior of the binuclear [{Ru(H2O)(bpz)}2(tpy2ph)](PF6)4 complex [bpz = 2,2'-bipyrazine; tpy2ph = 1,3-bis(4'-2,2':6',2''-terpyridin-4-yl)benzene] as compared with the structurally analogous derivative with 2,2'-bipyridine (bpy) instead of bpz. The differences were assigned to the stronger π-accepting character of bpz relative to bpy as the ancillary ligand. The expectation of a positive shift for the Ru-centered redox potentials was confirmed for the lower oxidation state species, but that trend was reversed in the formation of the high-valence catalytic active species as shown by a negative shift of 0.14 V for the potential of the [RuIV/V[double bond, length as m-dash]O] process. Moreover, DFT calculations indicated a significant decrease of about 15% on the spin density and oxyl character of the [RuV[double bond, length as m-dash]O]3+ fragment. The significantly lower kcat(O2) for the bpz system was attributed to these combined electronic effects.

Crystal structure and redox potentials of the tppz-bridged {RuCl(bpy)}+ dimer.

We report the structural and electrochemical characterization of the binuclear complex [µ-(C24H16N6){RuCl(C10H8N2)}2](PF6)2, which contains the bis-tridentate bridging ligand 2,3,5,6-tetra-kis-(pyridin-2-yl)pyrazine (tppz), the monodentate ligand Cl-, and the bidentate ligand 2,2'-bi-pyridine (bpy) {systematic name: µ-2,3,5,6-tetra-kis-(pyridin-2-yl)pyrazine-bis-[(2,2'-bi-pyridine)-chlorido-ru-thenium(II)] bis-(hexa-fluorido-phosphate)}. The complete [(bpy)(Cl)Ru(tppz)Ru(Cl)(bpy)]2+ dication is generated by crystallographic twofold symmetry; the tppz bridging ligand has a significantly twisted conformation, with an average angle of 42.4° between the mean planes of adjacent pyridyl rings. The metal-coordinated chloride ligands are in a trans configuration relative to each other across the {Ru(tppz)Ru} unit. The RuII ion exhibits a distorted octa-hedral geometry due to the restricted bite angle [160.6 (3)°] of the tppz ligand. For bpy, the bond lengths of the Ru-N bonds are 2.053 (8) and 2.090 (8) Å, with the shorter bond being opposite to Ru-Cl. For the tridentate tppz, the Ru-N distances involving the outer N atoms trans to each other are 2.069 (8) and 2.072 (9) Å, whereas the Ru-N bond involving the central N atom has the much shorter length of 1.939 (7) Šas a result of the geometric constraints and stronger π-acceptor ability of the pyrazine-centered bridge. The Ru-Cl distance is 2.407 (3) Šand the intra-molecular distance between Ru centers is 6.579 (4) Å. In the crystal, weak C-H⋯Cl and C-H⋯F inter-actions consolidate the packing.

Conjugation of Amphiphilic Proteins to Hydrophobic Ligands in Organic Solvent.

Protein-ligand conjugations are usually carried out in aqueous media in order to mimic the environment within which the conjugates will be used. In this work, we focus on the conjugation of amphiphilic variants of elastin-like polypeptide (ELP), short elastin (sEL), to poorly water-soluble compounds like OPPVs ( p-phenylenevinylene oligomers), triarylamines, and polypyridine-metal complexes. These conjugations are problematic when carried out in aqueous phase because hydrophobic ligands tend to avoid exposure to water, which in turn causes the ligand to self-aggregate and/or interact noncovalently with hydrophobic regions of the amphiphile. Ultimately, this behavior leads to low conjugation efficiency and contamination with strong noncovalent "conjugates". After exploring the solubility of sEL in various organic solvents, we have established an efficient conjugation methodology for obtaining covalent conjugates virtually free of contaminating noncovalent complexes. When conjugating carboxylated ligands to the amphiphile amines, we demonstrate that even when only one amine (the N-terminus) is present, its derivatization is 98% efficient. When conjugating amine moieties to the amphiphile carboxyls (a problematic configuration), protein multimerization is avoided, 98-100% of the protein is conjugated, and the unreacted ligand is recovered in pure form. Our syntheses occur in "one pot", and our purification procedure is a simple workup utilizing a combination of water and organic solvent extractions. This conjugation methodology might provide a solution to problems arising from solubility mismatch of protein and ligand, and it is likely to be widely applied for modification of recombinant amphiphiles used for drug delivery (PEG-antibodies, polymer-enzymes, food proteins), cell adhesion (collagen, hydrophobins), synthesis of nanostructures (peptides), and engineering of biocompatible optoelectronics (biological polymers), to cite a few.

A metallo-biopolymer conjugate of elastin-like polypeptide: photoluminescence enhancement in the coacervate microenvironment.

An optically active metallo-polymer assembly is demonstrated via conjugation of a genetically engineered elastin-like polypeptide (ELP) and a ruthenium(II) polypyridyl complex. By taking advantage of the phase transition of ELPs in water, photophysical properties of the resultant conjugate are investigated for both phases, below and above the critical transition temperature. Upon coacervation, the luminescence of the metallo-ELP is greatly enhanced as a consequence of local effects on the metal-ligand luminophore. These findings open a possibility to harness the temperature control of stimuli-responsive properties of biopolymers.

Evolution and characterization of a new reversibly photoswitching chromogenic protein, Dathail.

We report the engineering of a new reversibly switching chromogenic protein, Dathail. Dathail was evolved from the extremely thermostable fluorescent proteins thermal green protein (TGP) and eCGP123 using directed evolution and ratiometric sorting. Dathail has two spectrally distinct chromogenic states with low quantum yields, corresponding to absorbance in a ground state with a maximum at 389nm, and a photo-induced metastable state with a maximum at 497nm. In contrast to all previously described photoswitchable proteins, both spectral states of Dathail are non-fluorescent. The photo-induced chromogenic state of Dathail has a lifetime of ~50min at 293K and pH7.5 as measured by UV-Vis spectrophotometry, returning to the ground state through thermal relaxation. X-ray crystallography provided structural insights supporting a change in conformation and coordination in the chromophore pocket as being responsible for Dathail's photoswitching. Neutron crystallography, carried out for the first time on a protein from the green fluorescent protein family, showed a distribution of hydrogen atoms revealing protonation of the chromophore 4-hydroxybenzyl group in the ground state. The neutron structure also supports the hypothesis that the photo-induced proton transfer from the chromophore occurs through water-mediated proton relay into the bulk solvent. Beyond its spectroscopic curiosity, Dathail has several characteristics that are improvements for applications, including low background fluorescence, large spectral separation, rapid switching time, and the ability to switch many times. Therefore, Dathail is likely to be extremely useful in the quickly developing fields of imaging and biosensors, including photochromic Förster resonance energy transfer, high-resolution microscopy, and live tracking within the cell.

Stimuli-Responsive Genetically Engineered Polymer Hydrogel Demonstrates Emergent Optical Responses.

Biopolymer-based optical hydrogels represent an emerging class of materials with potential applications in biocompatible integrated optoelectronic devices, bioimaging applications, and stretchable/flexible photonics. We have synthesized stimuli-responsive three-dimensional hydrogels from genetically engineered elastin-like polymers (ELPs) and have loaded these hydrogels with an amine-containing p-phenylenevinylene oligomer (OPPV) derivative featuring highly tunable, environmentally sensitive optical properties. The composite ELP/OPPV hydrogels exhibit both pH- and temperature-dependent fluorescence emission, from which we have characterized a unique optical behavior that emerged from OPPV within the hydrogel environment. By systematic comparison with free OPPV in solution, our results suggest that this distinct behavior is due to local electronic effects arising from interactions between the hydrophobic ELP microenvironment and the nonprotonated OPPV species at pH 7 or higher.

Crystal structure of a mononuclear Ru(II) complex with a back-to-back terpyridine ligand: [RuCl(bpy)(tpy-tpy)](.).

We report the structural characterization of [6',6''-bis-(pyridin-2-yl)-2,2':4',4'':2'',2'''-quaterpyridine](2,2'-bi-pyridine)-chlorido-ruthenium(II) hexa-fluorido-phosphate, [RuCl(C10H8N2)(C30H20N6)]PF6, which contains the bidentate ligand 2,2'-bi-pyridine (bpy) and the tridendate ligand 6',6''-bis-(pyridin-2-yl)-2,2':4',4'':2'',2'''-quaterpyridine (tpy-tpy). The [RuCl(bpy)(tpy-tpy)](+) monocation has a distorted octa-hedral geometry at the central Ru(II) ion due to the restricted bite angle [159.32 (16)°] of the tridendate ligand. The Ru-bound tpy and bpy moieties are nearly planar and essentially perpendicular to each other with a dihedral angle of 89.78 (11)° between the least-squares planes. The lengths of the two Ru-N bonds for bpy are 2.028 (4) and 2.075 (4) Å, with the shorter bond being opposite to Ru-Cl. For tpy-tpy, the mean Ru-N distance involving the outer N atoms trans to each other is 2.053 (8) Å, whereas the length of the much shorter bond involving the central N atom is 1.936 (4) Å. The Ru-Cl distance is 2.3982 (16) Å. The free uncoordinated moiety of tpy-tpy adopts a trans,trans conformation about the inter-annular C-C bonds, with adjacent pyridyl rings being only approximately coplanar. The crystal packing shows significant π-π stacking inter-actions based on tpy-tpy. The crystal structure reported here is the first for a tpy-tpy complex of ruthenium.

A Hybrid DNA-Templated Gold Nanocluster For Enhanced Enzymatic Reduction of Oxygen.

We report the synthesis and characterization of a new DNA-templated gold nanocluster (AuNC) of ∼1 nm in diameter and possessing ∼7 Au atoms. When integrated with bilirubin oxidase (BOD) and single walled carbon nanotubes (SWNTs), the AuNC acts as an enhancer of electron transfer (ET) and lowers the overpotential of electrocatalytic oxygen reduction reaction (ORR) by ∼15 mV as compared to the enzyme alone. In addition, the presence of AuNC causes significant enhancements in the electrocatalytic current densities at the electrode. Control experiments show that such enhancement of ORR by the AuNC is specific to nanoclusters and not to plasmonic gold particles. Rotating ring disk electrode (RRDE) measurements confirm 4e(-) reduction of O2 to H2O with minimal production of H2O2, suggesting that the presence of AuNC does not perturb the mechanism of ORR catalyzed by the enzyme. This unique role of the AuNC as enhancer of ET at the enzyme-electrode interface makes it a potential candidate for the development of cathodes in enzymatic fuel cells, which often suffer from poor electronic communication between the electrode surface and the enzyme active site. Finally, the AuNC displays phosphorescence with large Stokes shift and microsecond lifetime.

A trinuclear oxo-chromium(III) complex containing the natural flavonoid primuletin: synthesis, characterization, and antiradical properties.

A new trinuclear oxo-centered chromium(III) complex with formula [Cr3O(CH3CO2)6(L)(H2O)2] (L = 5-hydroxyflavone, known as primuletin) was synthetized and characterized by ESI mass spectrometry, thermogravimetry, and 1H-NMR, UV-Vis, and FTIR spectroscopies. In agreement with the experimental results, DFT calculations indicated that the flavonoid ligand is coordinated to one of the three Cr(III) centers in an O,O-bidentate mode through the 5-hydroxy/4-keto groups. In a comparative study involving the uncoordinated primuletin and its corresponding complex, systematic reactions with the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) showed that antiradical activity increases upon complexation.

Thermal green protein, an extremely stable, nonaggregating fluorescent protein created by structure-guided surface engineering.

In this article, we describe the engineering and X-ray crystal structure of Thermal Green Protein (TGP), an extremely stable, highly soluble, non-aggregating green fluorescent protein. TGP is a soluble variant of the fluorescent protein eCGP123, which despite being highly stable, has proven to be aggregation-prone. The X-ray crystal structure of eCGP123, also determined within the context of this paper, was used to carry out rational surface engineering to improve its solubility, leading to TGP. The approach involved simultaneously eliminating crystal lattice contacts while increasing the overall negative charge of the protein. Despite intentional disruption of lattice contacts and introduction of high entropy glutamate side chains, TGP crystallized readily in a number of different conditions and the X-ray crystal structure of TGP was determined to 1.9 Å resolution. The structural reasons for the enhanced stability of TGP and eCGP123 are discussed. We demonstrate the utility of using TGP as a fusion partner in various assays and significantly, in amyloid assays in which the standard fluorescent protein, EGFP, is undesirable because of aberrant oligomerization.

Polythiophenes in biological applications.

Polythiophene and its derivatives have shown tremendous potential for interfacing electrically conducting polymers with biological applications. These semiconducting organic polymers are relatively soft, conduct electrons and ions, have low cytotoxicity, and can undergo facile chemical modifications. In addition, the reduction in electrical impedance of electrodes coated with polythiophenes may prove to be invaluable for a stable and permanent connection between devices and biological tissues. This review article focuses on the synthesis and some key applications of polythiophenes in multidisciplinary areas at the interface with biology. These polymers have shown tremendous potential in biological applications such as diagnostics, therapy, drug delivery, imaging, implant devices and artificial organs.

(4'-Ethynyl-2,2':6',2''-terpyridine)(2,2':6',2''-terpyridine)-ruthenium(II) bis-(hexa-fluoridophosphate) acetonitrile disolvate.

The title heteroleptic bis--terpyridine complex, [Ru(C(15)H(11)N(3))(C(17)H(11)N(3))](PF(6))(2)·2CH(3)CN, crystallized from an acetonitrile solution as a salt containing two hexa-fluoridophosphate counter-ions and two acetonitrile solvent mol-ecules. The Ru(II) atom has a distorted octa-hedral geometry due to the restricted bite angle [157.7 (3)°] of the two mer-arranged N,N',N''-tridendate ligands, viz. 2,2':6',2''-terpyridine (tpy) and 4'-ethynyl-2,2':6',2''-terpyridine (tpy'), which are essentially perpendicular to each other, with a dihedral angle of 87.75 (12)° between their terpyridyl planes. The rod-like acetyl-ene group lies in the same plane as its adjacent terpyridyl moiety, with a maximum deviation of only 0.071 (11) Šfrom coplanarity with the pyridine rings. The mean Ru-N bond length involving the outer N atoms trans to each other is 2.069 (6) Šat tpy and 2.070 (6) Šat tpy'. The Ru-N bond length involving the central N atom is 1.964 (6) Šat tpy and 1.967 (6) Šat tpy'. Two of the three counter anions were refined as half-occupied.

µ-2,3,5,6-Tetra-kis(pyridin-2-yl)pyrazine-bis-[(2,2':6',2''-terpyridine)-ruthenium(II)] tetra-kis-(hexa-fluoridophosphate) acetonitrile tetra-solvate.

In the title compound [Ru(2)(C(15)H(11)N(3))(2)(C(24)H(16)N(6))](PF(6))(4)·4CH(3)CN, two of the counter-ions and one of the solvent mol-ecules are disordered with occupancies for the major components between 0.57 (2) and 0.64 (1). The structure of the dinuclear tetracation exhibits significant distortion from planarity in the bridging 2,3,5,6-tetra-kis-(pyridin-2-yl)pyrazine (tppz) ligand, which has a saddle-like geometry with an average dihedral angle of 42.96 (18)° between adjacent pyridine rings. The metal-ligand coordination environment is nearly equivalent for the two Ru(II) atoms, which have a distorted octa-hedral geometry due to the restricted bite angle [157.57 (13)-159.28 (12)°] of their two mer-arranged tridendate ligands [2,2':6',2''-terpyridine (tpy) and tppz] orthogonal to each other. At the peripheral tpy ligands, the average Ru-N bond distance is 2.072 (4) Šfor the outer N atoms trans to each other (N(outer)) and 1.984 (1) Šfor the central N atoms (N(central)). At the bridging tppz ligand, the average metal-ligand distances are significantly shorter [2.058 (4) Šfor Ru-N(outer) and 1.965 (1) Šfor Ru-N(central)] as a result of both the geometric constraints and the stronger π-acceptor ability of the pyrazine-centered bridge. The dihedral angle between the two tpy planes is 27.11 (6)°. The intra-molecular linear distance between the two Ru atoms is 6.6102 (7) Å.

(2,2'-Bi-pyridine)-chlorido-[diethyl (2,2':6',2''-terpyridin-4-yl)phospho-nate]ruthenium(II) hexa-fluorido-phosphate aceto-nitrile/water solvate.

The cationic complex in the title compound, [RuCl(C10H8N2)(C19H20N3O3P)]PF6·0.83CH3CN·0.17H2O, is a water-oxidation precatalyst functionalized for TiO2 attachment via terpyridine phospho-nate. The The Ru(II) atom in the complex has a distorted octa-hedral geometry due to the restricted bite angle [159.50 (18)°] of the terpyridyl ligand. The dihedral angle between the least-squares planes of the terpyridyl and bipyridyl moieties is 86.04 (14)°. The mean Ru-N bond length for bi-pyridine is 2.064 (5) Å, with the bond opposite to Ru-Cl being 0.068 Šshorter. For the substituted terpyridine, the mean Ru-N distance involving the outer N atoms trans to each other is 2.057 (6) Å, whereas the bond length involving the central N atom is 1.944 (5) Å. The Ru-Cl distance is 2.4073 (15) Å. The P atom of the phospho-nate group lies in the same plane as its adjacent pyridyl ring, with the ordinary character of the bond between P and Ctpy [1.801 (6) Å] allowing for free rotation of the terpyridine substituent around the P-Ctpy axis. The aceto-nitrile solvent mol-ecule was refined to be disordered with two water mol-ecules; occupancies for the acetontrile and water mol-ecules were 0.831 (9) and 0.169 (9), respectively. Also disordered was the PF6 (-) counter-ion (over three positions) and one of the eth-oxy substituents (with two positions). The crystal structure shows significant intra- and inter-molecular H⋯X contacts, especially some involving the Cl(-) ligand.

A DNA-templated fluorescent silver nanocluster with enhanced stability.

We report the discovery of a DNA sequence that templates a highly stable fluorescent silver nanocluster. In contrast to other DNA templated silver nanoclusters that have a relatively short shelf-life, the fluorescent species templated in this new DNA sequence retains significant fluorescence for at least a year. Moreover, this new silver nanocluster possesses low cellular toxicity and enhanced thermal, oxidative, and chemical stability.

Electronic structure of the water oxidation catalyst cis,cis-[(bpy)2(H2O)Ru(III)ORu(III)(OH2)(bpy)2]4+, the blue dimer.

The first designed molecular catalyst for water oxidation is the "blue dimer", cis,cis-[(bpy)(2)(H(2)O)Ru(III)ORu(III)(OH(2))(bpy)(2)](4+). Although there is experimental evidence for extensive electronic coupling across the µ-oxo bridge, results of earlier DFT and CASSCF calculations provide a model with magnetic interactions of weak to moderately coupled Ru(III) ions across the µ-oxo bridge. We present the results of a comprehensive experimental investigation, combined with DFT calculations. The experiments demonstrate both that there is strong electronic coupling in the blue dimer and that its effects are profound. Experimental evidence has been obtained from molecular structures and key bond distances by XRD, electrochemically measured comproportionation constants for mixed-valence equilibria, temperature-dependent magnetism, chemical properties (solvent exchange, redox potentials, and pK(a) values), XPS binding energies, analysis of excitation-dependent resonance Raman profiles, and DFT analysis of electronic absorption spectra. The spectrum can be assigned based on a singlet ground state with specific hydrogen-bonding interactions with solvent molecules included. The results are in good agreement with available experimental data. The DFT analysis provides assignments for characteristic absorption bands in the near-IR and visible regions. Bridge-based dπ → dπ* and interconfiguration transitions at Ru(III) appear in the near-IR and MLCT and LMCT transitions in the visible. Reasonable values are also provided by DFT analysis for experimentally observed bond distances and redox potentials. The observed temperature-dependent magnetism of the blue dimer is consistent with a delocalized, diamagnetic singlet state (dπ(1)*)(2) with a low-lying, paramagnetic triplet state (dπ(1)*)(1)(dπ(2)*)(1). Systematic structural-magnetic-IR correlations are observed between ν(sym)(RuORu) and ν(asym)(RuORu) vibrational energies and magnetic properties in a series of ruthenium-based, µ-oxo-bridged complexes. Consistent with the DFT electronic structure model, bending along the Ru-O-Ru axis arises from a Jahn-Teller distortion with ∠Ru-O-Ru dictated by the distortion and electron-electron repulsion.

Catalytic photooxidation of alcohols by an unsymmetrical tetra(pyridyl)pyrazine-bridged dinuclear Ru complex.

The dinuclear complexes [(tpy)Ru(tppz)Ru(bpy)(L)](n+) (where L is Cl(-) or H(2)O, tpy and bpy are the terminal ligands 2,2':6',2''-terpyridine and 2,2'-bipyridine, and tppz is the bridging backbone 2,3,5,6-tetrakis(2-pyridyl)pyrazine) were prepared and structurally and electronically characterized. The mononuclear complexes [(tpy)Ru(tppz)](2+) and [(tppz)Ru(bpy)(L)](m+) were also prepared and studied for comparison. The proton-coupled, multi-electron photooxidation reactivity of the aquo dinuclear species was shown through the photocatalytic dehydrogenation of a series of primary and secondary alcohols. Under simulated solar irradiation and in the presence of a sacrificial electron acceptor, the photoactivated chromophore-catalyst complex (in aqueous solutions at room temperature and ambient pressure conditions) can perform the visible-light-driven conversion of aliphatic and benzylic alcohols into the corresponding carbonyl products (i.e., aldehydes or ketones) with 100% product selectivity and several tens of turnover cycles, as probed by NMR spectroscopy and gas chromatography. Moreover, for aliphatic substrates, the activity of the photocatalyst was found to be highly selective toward secondary alcohols, with no significant product formed from primary alcohols. Comparison of the activity of this tppz-bridged complex with that of the analogue containing a back-to-back terpyridine bridge (tpy-tpy, i.e., 6',6''-bis(2-pyridyl)-2,2':4',4'':2'',2'''-quaterpyridine) demonstrated that the latter is a superior photocatalyst toward the oxidation of alcohols. The much stronger electronic coupling with significant delocalization across the strongly electron-accepting tppz bridge facilitates charge trapping between the chromophore and catalyst centers and therefore is presumably responsible for the decreased catalytic performance.