Introduction of sulfhydryl groups and disulfide linkage to mungbean 8Sα globulin and effects on physicochemical and functional properties

Sulfhydryl groups and disulfide bond were introduced in mungbean's major storage protein, 8Sα globulin, by protein engineering to improve structural stability and functional properties. Five modified proteins or mutants (F59C, I99C, A213C, one free sulfhydryl group; I99C/A213, one disulfide bridge; F59C/I99C/A213C, one free sulfhydryl group and one disulfide linkage) were expressed in Escherichia coli at a yield similar to that of the unmodified protein or wild type (WT) in soluble form (38%). The number of introduced groups in the mutants was confirmed by Ellman analysis. Mutant and WT proteins exhibited similar elution patterns on gel filtration indicating their trimeric native conformation. Mutants had 2 to 3.8 °C higher T_m values than WT and were digested by chymotrypsin at 52–58% in 60 min but exhibited different digestion patterns. All mutants showed greater hardness of heat-induced gels than WT, especially I99C/A213C and F59C/I99C/A213C. Results indicate the improved structural stability of the modified 8Sα globulin.     

Mammalian Cytochrome P450-Dependent Metabolism of Polychlorinated Dibenzo-p-dioxins and Coplanar Polychlorinated Biphenyls

Polychlorinated dibenzo-p-dioxins (PCDDs) and coplanar polychlorinated biphenyls (PCBs) contribute to dioxin toxicity in humans and wildlife after bioaccumulation through the food chain from the environment. The authors examined human and rat cytochrome P450 (CYP)-dependent metabolism of PCDDs and PCBs. A number of human CYP isoforms belonging to the CYP1 and CYP2 families showed remarkable activities toward low-chlorinated PCDDs. In particular, human CYP1A1, CYP1A2, and CYP1B1 showed high activities toward monoCDDs, diCDDs, and triCDDs but no detectable activity toward 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-tetraCDD). Large amino acids located at putative substrate-recognition sites and the F-G loop in rat CYP1A1 contributed to the successful metabolism of 2,3,7,8-tetraCDD. Rat, but not human, CYP1A1 metabolized 3,3'',4,4'',5-pentachlorobiphenyl (CB126) to two hydroxylated metabolites. These metabolites are probably less toxic than is CB126, due to their higher solubility. Homology models of human and rat CYP1A1s and CB126 docking studies indicated that two amino acid differences in the CB126-binding cavity were important for CB126 metabolism. In this review, the importance of CYPs in the metabolism of dioxins and PCBs in mammals and the species-based differences between humans and rats are described. In addition, the authors reveal the molecular mechanism behind the binding modes of dioxins and PCBs in the heme pocket of CYPs.     

Improvement of a Measurement System for Surface Loss Rate of Ozone

Ozone molecules present in high-purity oxygen as a feed gas interact with surfaces and oxygen molecules in an enclosed container. Therefore, some of the ozone molecules are destroyed and the density of ozone decreases with time. An experimental system has been constructed to monitor the temporal decrease in ozone density in a container based on the HgI 254 nm photoabsorption method. Our investigation is focused on the dependence of the effective lifetime of ozone on the wall material at various gas pressures. During the experiments, distortion of the measured decay curves often caused by instabilities in the mercury lamp intensity. We thus attempted to build a simple setup that eliminates the effect of long-term intensity drift. This setup is based on light source intensity monitoring by a separate photodetector and data correction software. This article describes the setup of the experimental apparatus, the results including some preliminary experiments and the temporal variation of the surface loss rate of ozone by repeated measurement along with inspections of the wall surface by Auger electron spectroscopy.     

A control law for vehicle merging inspired by dragonfly behavior

Autonomous control of vehicles has recently attracted considerable attention. In this sense, vehicle merging has become an important topic in this field of research. However, in conventional studies, the controlled vehicle must calculate the movement of other surrounding vehicles to complete the merge, requiring high computational costs. In this paper, we focus on dragonfly behavior to solve this issue. Indeed, insects can behave adaptively in the complex real world in spite of the limited size of their brains. They reduce the computational requirements of their brain by relying on different properties of their surroundings, basing their intelligent behaviors on simple strategies. The behavior of a dragonfly when chasing a prey is an example of these strategies. In this study, we address the vehicle merging maneuver by applying dragonfly’s strategies to control the movement of the merging vehicle. We propose a simple control method inspired by the aforementioned strategies and, finally, we present simulation results that were conducted to demonstrate the effectiveness of this method.     

Euphorbia tirucalli β-Amyrin Synthase: Critical Roles of Steric Sizes at Val483 and Met729 and the CH–π Interaction between Val483 and Trp534 for Catalytic Action

The functions of Val483, Trp534, and Met729 in Euphorbia tirucalli β-amyrin synthase were revealed by comparing the enzyme activities of site-directed mutants against that of the wild type. The Gly and Ala variants with a smaller bulk size at position 483 predominantly afforded monocyclic camelliol C, which suggested that the orientation of the (3S)-2,3-oxidosqualene substrate was not appropriately arranged in the reaction cavity as a result of the decreased bulk size, leading to failure of its normal folding into the chair–chair–chair–boat–boat conformation. The Ile variant, with a somewhat larger bulk, afforded β-amyrin as the dominant product. Intriguingly, various variants of Trp534 exhibited significantly decreased enzymatic activities and provided no aberrantly cyclized products, although the aromatic Phe and Tyr residues were incorporated and the steric sizes of the aliphatic residues were altered. Therefore, the Trp534 residue does not stabilize the transient cation through a cation–π interaction. Furthermore, the Trp residue, with the largest steric bulk among all natural amino acids, is essential for high enzymatic activity. Robust CH–π complexation between the Val483 and Trp534 residues is proposed herein. Altering the steric bulk at the Met729 position afforded the pentacyclic skeletons. Thus, Met729 is positioned at the E-ring formation site. More detailed insights into the functions of the Val483, Trp534, and Met729 residues are provided by homology modeling.     

Chiral lewis acid-catalyzed enantioselective michael reactions in water

Asymmetric Michael reactions of ß-ketoesters with enones were effectively catalyzed by complexes of silver salts and BINAP derivatives in water. The reactions have proved to give Michael adducts in high yields with high enantioselectivities. These reactions are the first example of Lewis acid-catalyzed asymmetric Michael reactions in water.     

Regional income disparities in an OLG structure

This study examines a two-region OLG mode, in which parents educate their children and choose their location freely; that is, the population distribution and each worker’s productivity are determined endogenously. Owing to the setting of both agglomeration economies in wage and agglomeration diseconomies in utility and thereby different consumption patterns between regions, the present model represents a generation of inequality in educational level between regions even when households with equivalent education are given. Incentive for interregional segregation by educational level is also assumed and inequality is persistent in the result. This model demonstrates a development process starting with a low-developed initial state that represents population concentration accompanied with human capital growth and expansion of interregional inequalities. In addition, the effects of policies on long-run equilibria are addressed along with lock-in effects against these policies.