Tomohiko Inomata

Creation of a type 1 blue copper site within a de novo coiled-coil protein scaffold.

Type 1 blue copper proteins uniquely coordinate Cu(2+) in a trigonal planar geometry, formed by three strong equatorial ligands, His, His, and Cys, in the protein. We designed a stable Cu(2+) coordination scaffold composed of a four-stranded α-helical coiled-coil structure. Two His residues and one Cys residue were situated to form the trigonal planar geometry and to coordinate the Cu(2+) in the hydrophobic core of the scaffold. The protein bound Cu(2+), displayed a blue color, and exhibited UV-vis spectra with a maximum of 602-616 nm, arising from the thiolate-Cu(2+) ligand to metal charge transfer, depending on the exogenous axial ligand, Cl(-) or HPO(4)(2-). The protein-Cu(2+) complex also showed unresolved small A(∥) values in the electron paramagnetic resonance (EPR) spectral analysis and a 328 mV (vs normal hydrogen electrode, NHE) redox potential with a fast electron reaction rate. The X-ray absorption spectrum revealed that the Cu(2+) coordination environment was identical to that found in natural type 1 blue copper proteins. The extended X-ray absorption fine structure (EXAFS) analysis of the protein showed two typical Cu-N(His) at around 1.9-2.0 Å, Cu-S(Cys) at 2.3 Å, and a long Cu-Cl at a 2.66 Å, which are also characteristic of the natural type 1 blue copper proteins.

A ligand substitution reaction of oxo-centred triruthenium complexes assembled as monolayers on gold electrodes

A terminal ligand-substitution reaction at triruthenium redox centres within monolayer films is investigated. For this purpose, a new redox-active trinuclear ruthenium complex containing one carbonyl ligand at a terminal site, [Ru3(O)(CH3COO)6(CO)(mpy)(C10PY)] (mpy = 4-methylpyridine, C10PY = {(NC5H4)CH2NHC(O)(CH2)10S–}2) 1, has been chemically adsorbed onto a gold electrode surface with a disulfide-alkyl spacer C10PY. Densely packed monolayers of complex 1 on Au electrodes (surface coverage, Γ(Ru3) = 1.8 × 10−10 mol cm−2) display the {RuII–CO}RuIIIRuIII/{RuIII–CO}RuIIIRuIII couple at E1/2 = + 0.61 V s. Ag–AgCl in a 0.1 mol dm−3 HClO4 aqueous solution. Upon electrochemical oxidation of the adsorbed complex at + 0.80 V, where the triruthenium carbonyl complex is in the oxidized form {RuIII–CO}RuIIIRuIII, the coordinated CO is replaced by an aqua ligand present in the bulk phase, which is followed by cyclic voltammetry at appropriate electrolysis time intervals. During 8000 s of electrolysis, ca. 67% of the initially assembled carbonyl complexes are converted to the triruthenium aqua complexes, while ca. 25% desorbed from the electrode surface.

Electrocatalytic Hydrogen Production by a Nickel(II) Complex with a Phosphinopyridyl Ligand

A novel nickel(II) complex [Ni(L)2 Cl]Cl with a bidentate phosphinopyridyl ligand 6-((diphenylphosphino)methyl)pyridin-2-amine (L) was synthesized as a metal-complex catalyst for hydrogen production from protons. The ligand can stabilize a low Ni oxidation state and has an amine base as a proton transfer site. The X-ray structure analysis revealed a distorted square-pyramidal Ni(II)  complex with two bidentate L ligands in a trans arrangement in the equatorial plane and a chloride anion at the apex. Electrochemical measurements with the Ni(II) complex in MeCN indicate a higher rate of hydrogen production under weak acid conditions using acetic acid as the proton source. The catalytic current increases with the stepwise addition of protons, and the turnover frequency is 8400 s(-1) in 0.1 m [NBu4 ][ClO4 ]/MeCN in the presence of acetic acid (290 equiv) at an overpotential of circa 590 mV.

The effect of the side chain length of Asp and Glu on coordination structure of Cu2+ in a de novo designed protein

Metal ions in proteins are important not only for the formation of the proper structures but also for various biological activities. For biological functions such as hydrolysis and oxidation, metal ions often adopt unusual coordination structures. We constructed a stable scaffold for metal binding to create distorted metal coordination structures. A stable four stranded alpha-helical coiled-coil structure was used as the scaffold, and the metal binding site was in the cavity created at the center of the structure. Two His residues and one Asp or Glu residue were used to coordinate the metal ions, AM2D and AM2E, respectively. Cu(2+) bound to AM2D with an equatorial planar coordination structure with two His, one Asp, and H(2)O as detected by electron spin resonance and UV spectral analyzes. On the other hand, Cu(2+) had a slightly distorted square planar structure when it bound two His and Glu in AM2E, due to the longer side-chain of the Glu residue as compared to the Asp residue. Computational analysis also supported the distorted coordination structure of Cu(2+) in AM2E. This construct should be useful to create various coordinations of metal ions for catalytic functions.

Self-assembled monolayer electrode of a diiron complex with a phenoxo-based dinucleating ligand: observation of molecular oxygen adsorption/desorption in aqueous media

The phenoxo-based dinucleating ligand, 2,6-bis[bis(6-pivalamido-2-pyridylmethyl)amino-methyl]-4-aminophenol (1), and its Fe2(II) complex, [Fe2(II)(1)(PhCOO)2](CF3SO3) (2), were prepared and 2 deposited on the Au surface (2/Au) is much more stable than in solution and exhibits redox behavior in aqueous media as well as reversible adsorption/desorption of oxygen at room temperature.

N2 Activation by an Iron Complex with a Strong Electron-Donating Iminophosphorane Ligand

A new tridentate cyclopentane-bridged iminophosphorane ligand, N-(2-diisopropylphosphinophenyl)-P,P-diisopropyl-P-(2-(2,6-diisopropylphenylamido)cyclopent-1-enyl)phosphoranimine (NpNPiPr), was synthesized and used in the preparation of a diiron dinitrogen complex. The reaction of the iron complex FeBr(NpNPiPr) with KC8 under dinitrogen yielded the dinuclear dinitrogen Fe complex [Fe(NpNPiPr)]2(μ-N2), which was characterized by X-ray analysis and resonance Raman and NMR spectroscopies. The X-ray analysis revealed a diiron complex bridged by the dinitrogen molecule, with each metal center coordinated by an NpNPiPr ligand and dinitrogen in a trigonal-monopyramidal geometry. The N–N bond length is 1.184(6) Å, and resonance Raman spectra indicate that the N–N stretching mode ν(14N2/15N2) is 1755/1700 cm–1. The magnetic moment of [Fe(NpNPiPr)]2(μ-N2) in benzene-d6 solution, as measured by 1H NMR spectroscopy by the Evans method, is 6.91μB (S = 3). The Mössbauer spectrum at 78 K showed δ = 0.73 mm/s and ΔEQ = 1.83 mm/s. These findings suggest that the iron ions are divalent with a high-spin configuration and that the N2 molecule has (N═N)2– character. Density functional theory calculations performed on [Fe(NpNPiPr)]2(μ-N2) also suggested that the iron is in a high-spin divalent state and that the coordinated dinitrogen molecule is effectively activated by π back-donation from the two iron ions (dπ) to the dinitrogen molecule (πx* and πy*). This is supported by cooperation between a large negative charge on the iminophosphorane ligand and strong electron donation and effective orbital overlap between the iron dπ orbitals and N2 π* orbitals supplied by the phosphine ligand.

Adsorption Behavior of Microbes on a QCM Chip Modified with an Artificial Siderophore–Fe3+ Complex

Three hydroxamate-type artificial siderophores with terminal NH(2) groups, tris[2-{3-(N-acyl-N-hydroxamino)propylamido}propyl]aminomethane (1-3, acyl-R group = Me, Et, and Ph, respectively), and their Fe(3+) complexes, 4-6, were prepared. The stability constant (log β) of 4 was estimated to be about 31 by its EDTA titration. The biological activities of 4-6 for Microbacterium flavescens, which is a hydroxamate-type siderophore, auxotrophic gram-positive microbe, clearly indicated that they permeated the cell membrane depending on their terminal bulky acyl-R groups. These artificial siderophore complexes, 4-6, were modified on Au electrode surfaces with the terminal NH(2) group (4-6/Au). The surface modification of 4-6 was confirmed by several electrochemical measurements. The quartz crystal microbalance (QCM) chips were also modified with 4-6. Microbe adsorption measurements using these modified QCM chips for M. flavescens, Pseudomonas putida, and Eschrichia coli were performed. The QCM chips have the ability to adsorb microbes selectively as a result of the differences in the interactions between the structures of Fe(3+)-artificial siderophore complexes and their receptors or binding proteins within the cell membrane.

Gold nanocluster confined within a cage: template-directed formation of a hexaporphyrin cage and its confinement capability

A novel container complex in which a 1.4 nm gold cluster is confined within a hexaporphyrin cage was synthesized; the cage showed notable confinement capability for the cluster core, but allowed the interpenetration of small molecules into the interstitial space.

Adsorption and detection of Escherichia coli using an Au substrate modified with a catecholate-type artificial siderophore–Fe3+ complex

A catecholate-type artificial siderophore with a terminal-NH2 group (1) and its Fe(3+) complex (2) were prepared. Siderophore 1 was characterized by (1)H NMR, FT-IR, and ESI-TOF MS spectroscopy. The corresponding Fe(3+) complex 2 was obtained by reaction of 1 with Fe(acac)3. The absorption band at 500 nm (ε = 4670 M(-1) cm(-1) at pH 7.0) of the electronic absorption spectrum of 2 is assignable as the LMCT (O(catecholate) → Fe(3+)) absorption band. This band indicates the formation of the Fe(3+) complex of 1. The biological activity of 2 with respect to Escherichia coli was clearly confirmed by observing that it permeates into the cell membrane. The self-assembled monolayer of 2 on an Au substrate, 2/Au, was prepared and its preparation was confirmed by FT-IR reflection-absorption spectroscopy (IR-RAS) and cyclic voltammetry (CV). Furthermore, a quartz crystal microbalance (QCM) chip modified with 2 effectively adsorbed E. coli. M. flavescens, an organism which is incapable of synthesizing siderophores and must therefore use exogenous hydroxamate-type siderophores for growth, did not adsorb on 2/Au. In contrast, E. coli did not adsorb on the hydroxamate-type artificial siderophore-Fe(3+) complex (3)-modified Au substrate, 3/Au. These results provide preliminary evidence that microbes recognized Fe(3+) ion-bound siderophores on the surface. The detection limit of 2/Au was ∼10(4) CFU mL(-1).

CCDC 1588610: Experimental Crystal Structure Determination

Related Article: Takuma Yano, Yuko Wasada-Tsutsui, Tomohiro Ikeda, Tomonori Shibayama, Yuji Kajita, Tomohiko Inomata, Yasuhiro Funahashi, Tomohiro Ozawa, Hideki Masuda|2018|Inorg.Chem.|57|4277|doi:10.1021/acs.inorgchem.6b02324