Live‐Cell Protein Sulfonylation Based on Proximity‐driven N‐Sulfonyl Pyridone Chemistry

The development of bioorthogonal approaches for labeling of endogenous proteins under the multimolecular crowding conditions of live cells is highly desirable for the analysis and engineering of proteins without using genetic manipulation. N‐Sulfonyl pyridone (SP) is reported as a new reactive group for protein sulfonylation. The ligand‐directed SP chemistry was able to modify not only purified proteins in vitro, but also endogenous ones on the surface of and inside live cells selectively and rapidly, which allowed to convert endogenous proteins to FRET‐based biosensors in situ.     

A Peptidyl‐Transesterifying Type I Thioesterase in Salinamide Biosynthesis

Salinamide A belongs to a rare class of bicyclic depsipeptide antibiotics in which the installation of a (4‐methylhexa‐2,4‐dienoyl)glycine handle across a hexadepsipeptide core contributes to its chemical complexity and biological properties. Herein, we report the genetic and biochemical basis for salinamide construction in the marine bacterium Streptomyces sp. CNB‐091, which involves a novel intermolecular transesterification reaction catalyzed by a type I thioesterase. Heterologous expression studies revealed the central role of the nonribosomal peptide synthetase Sln9 in constructing and installing the distinctive acylglycine “basket handle” of salinamide. Biochemical characterization of the Sln9 thioesterase domain established that transesterification of the serine residue of desmethylsalinamide E with acylated glycyl thioesters yields desmethylsalinamide C.     

Relationship between size and surface modification of silica particles and enhancement and suppression of inflammatory cytokine production by lipopolysaccharide- or peptidoglycan-stimulated RAW264.7 macrophages

Although nanomaterials are used in an increasing number of commodities, the relationships between their immunotoxicity and physicochemical properties such as size or surface characteristics are not fully understood. Here we demonstrated that pretreatment with amorphous silica particles (SPs) of various sizes (diameters of 10–1000 nm), with or without amine surface modification, significantly decreased interleukin 6 production by RAW264.7 macrophages following lipopolysaccharide or peptidoglycan stimulation. Furthermore, nanosized, but not microsized, SPs significantly enhanced tumor necrosis factor-α production in macrophages stimulated with lipopolysaccharide. This altered cytokine response was distinct from the inflammatory responses induced by treatment with the SPs alone. Additionally, the uptake of SPs into macrophages by phagocytosis was found to be crucial for the suppression of macrophage immune response to occur, irrespective of particle size or surface modification. Together, these results suggest that SPs may not only increase susceptibility to microbial infection, but that they may also be potentially effective immunosuppressants.     

Synthesis and Disassembly/Reassembly of Giant Ring‐Shaped Polyoxotungstate Oligomers

The disassembly and reassembly of giant molecules are essential processes in controlling the structure and function of biological and artificial systems. In this work, the disassembly and reassembly of a giant ring‐shaped polyoxometalate (POM) without isomerization of the monomeric units is reported. The reaction of a hexavacant lacunary POM that is soluble in organic solvents, [P₂W₁₂O₄₈]^14?, with manganese cations gave the giant ring‐shaped POM [{γ‐P₂W₁₂O₄₈Mn₄(C₅H₇O₂)₂(CH₃CO₂)}₆]^42?. This POM is a hexamer of manganese‐substituted {P₂W₁₂O₄₈Mn₄} units, and its inner cavity was larger than any of those previously reported for ring‐shaped polyoxotungstates. It was disassembled into monomeric units in acetonitrile, and the removal of the capping organic ligands on the manganese cations led to reassembly into a tetrameric ring‐shaped POM, [{γ‐P₂W₁₂O₄₈Mn₄(H₂O)₆}₄(H₂O)₄]^24?.     

Local State and Sector Theory in Local Quantum Physics

We define a new concept of local states in the framework of algebraic quantum field theory (AQFT). Local states are a natural generalization of states and give a clear vision of localization in the context of QFT. In terms of them, we can find a condition from which follows automatically the famous DHR selection criterion in DHR-DR theory. As a result, we can understand the condition as consequences of physically natural state preparations in vacuum backgrounds. Furthermore, a theory of orthogonal decomposition of completely positive (CP) maps is developed. It unifies a theory of orthogonal decomposition of states and order structure theory of CP maps. Using it, localized version of sectors is formulated, which gives sector theory for local states with respect to general reference representations.     

Dinitrogen Activation and Addition to Unsaturated C−E (E=C,N, O,S) Bonds Mediated by Transition Metal Complexes

Dinitrogen (N₂) activation and functionalization is of fundamental interest and practical importance. This review focuses on N₂ activation and addition to unsaturated substrates, including carbon monoxide, carbon dioxide, heteroallenes, aldehydes, ketones, acid halides, nitriles, alkynes, and allenes, mediated by transition metal complexes, which afforded a variety of N−C bond formation products. Emphases are placed on the reaction modes and mechanisms. We hope that this work would stimulate further explorations in this challenging field.     

A Gd3+-based magnetic resonance imaging contrast agent sensitive to beta-galactosidase activity utilizing a receptor-induced magnetization enhancement (RIME) phenomenon

Magnetic resonance imaging (MRI) permits noninvasive three-dimensional imaging of opaque organisms. Gadolinium (Gd(3+)) complexes have become important imaging tools as MRI contrast agents for MRI studies, though most of them are nonspecific and report solely on anatomy. Recently, MRI contrast agents have been reported whose ability to relax water protons is triggered or greatly enhanced by recognition of a particular biomolecule. This new class of MRI contrast agents could open up the possibility of reporting on the physiological state or metabolic activity deep within living specimens. One possible strategy for this purpose is to utilize the increase in the longitudinal water proton r(1) relaxivity that occurs upon slowing the molecular rotation of a small paramagnetic complex, a phenomenon which is known as receptor-induced magnetization enhancement (RIME), by either binding to a macromolecule or polymerization of the agent itself. Here we describe the design and synthesis of a novel beta-galactosidase-activated MRI contrast agent, the Gd(3+) complex [Gd-5], by using the RIME approach. beta-Galactosidase is commonly used as a marker gene to monitor gene expression. This newly synthesized compound exhibited a 57% increase in the r(1) relaxivity in phosphate-buffered saline (PBS) with 4.5% w/v human serum albumin (HSA) in the presence of beta-galactosidase. Detailed investigations revealed that RIME is the dominant factor in this increase of the observed r(1) relaxivity, based on analysis of Gd(3+) complexes [Gd-5] and [Gd-8], which is generated from [Gd-5] by the activity of beta-galactosidase, and spectroscopic analysis of their corresponding Tb(3+) complexes, [Tb-5] and [Tb-8].     

Selective isolation of N-terminal peptides from proteins and their de novo sequencing by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry without regard to unblocking or blocking of N-terminal amino acids

We have developed a new method to determine the N-terminal amino acid sequences of proteins, regardless of whether their N-termini are modified. This method consists of the following five steps: (1) reduction, S-alkylation and guanidination for targeted proteins; (2) coupling of sulfo-NHS-SS-biotin to N(alpha)-amino groups of proteins; (3) digestion of the modified proteins by an appropriate protease followed by oxidation with performic acid; (4) specific isolation of N-terminal peptides from digests using DITC resins; (5) de novo sequence analysis of the N-terminal peptides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) using the CAF (chemically assisted fragmentation) method or tandem mass spectrometric (MS/MS) analysis according to unblocked or blocked peptides, respectively. By employing DITC resins instead of avidin resins used in our previous method (Yamaguchi et al., Rapid Commun. Mass Spectrom. 2007; 21: 3329), it has been possible to isolate selectively N-terminal peptides from proteins regardless of modification of N-terminal amino acids. Here we propose a universal method for N-terminal sequence analysis of proteins.     

DNA liquid-crystalline gel as adsorbent of carcinogenic agent

DNA liquid crystalline gel (LCG) has been newly prepared by a dialysis of concentrated DNA solutions into concentrated metal cation solutions. The condition for forming DNA LCG is examined by means of the insolubilization reaction. The shrinking ratio and the ratio of the thickness of LCG layer, delta, and the diameter of the dialysis tube, d0, do not depend on d0. The adsorption of one of carcinogenic agents, acridine orange is demonstrated. From the experimental results, the mechanism for forming DNALCG is discussed.     

Estimation of Silica Species’ Concentrations in Lithium and Magnesium Chloride Solutions from the Peak Intensities Observed by FAB-MS

The concentrations of dissolved silica species in electrolyte solutions were derived from the relative intensities of silica species, obtained from FAB-MS measurements (fast atom bombardment mass spectrometry), and the total concentration of dissolved silica. Generally, silica species in aqueous solutions form various complexes with cations such as sodium (Na⁺) or calcium (Ca²⁺), and it has been difficult to determine the concentration of each species. From the observed results from FAB-MS, the chemical species of silica dissolved in lithium chloride (LiCl) and magnesium chloride (MgCl₂) solutions do not include complexes with these cations, and thus Li⁺ and Mg²⁺ do not replace protons of the silanol groups in silica. Therefore, in LiCl and MgCl₂ solutions, all of the simple structures of silicate species can be identified. The concentration of each silica species was estimated on the basis of its mass spectra peak intensities and the total concentration of silica as determined by colorimetry. This study yields the concentration of each silica species within small errors, whereas conventional methods (such as ²⁹Si-NMR) have not yielded the concentrations of individual silica species. From these results, dimers and cyclic tetramers are concluded to be the main species in silica solutions with concentrations of at most 0.1 to 0.2 μmol⋅dm⁻³. This tendency should also occur in NaCl and CaCl₂ solutions, which are major electrolytes in natural waters.