The Emi2 Protein of Saccharomyces cerevisiae is a Hexokinase Expressed under Glucose Limitation

Hexokinases catalyze glucose phosphorylation at the first step in glycolysis in eukaryotes. In the budding yeast Saccharomyces cerevisiae , three enzymes for glucose phosphorylation have long been known: Hxk1, Hxk2, and Glk1. In this study, we focus on Emi2, a previously uncharacterized hexokinase-like protein of S. cerevisiae . Our data show that the recombinant Emi2 protein (rEmi2), expressed in Escherichia coli , possesses glucose-phosphorylating activity in the presence of ATP and Mg ²⁺ . It was also found that rEmi2 phosphorylates not only glucose but also fructose, mannose and glucosamine in vitro . In addition, we examined changes in the level of endogenous Emi2 protein in S. cerevisiae in the presence or absence of glucose and a non-fermentable carbon source. We found that the expression of Emi2 protein is tightly suppressed during proliferation in high glucose, while it is strongly upregulated in response to glucose limitation and the presence of a non-fermentable carbon source. Our data suggest that the expression of the endogenous Emi2 protein in S. cerevisiae is regulated under the control of Hxk2 in response to glucose availability in the environment.     

Oxidation of doped europium in BaMgAl10O17 by annealing studied by x‐ray‐absorption fine‐structure measurements

Abstract— We studied the influence of annealing in air on doped europium in BaMgAl₁₀O₁₇ by performing x‐ray absorption fine‐structure measurements. We determined the oxidation of doped divalent europium by annealing in air at over 500°C. The interatomic distance between the europium and the surrounding oxygen atoms was compressed by oxidation. It also appears that the oxidation process of europium is determined by the diffusion of oxygen into BaMgAl₁₀O₁₇.     

Effect of starting particle size on hot-pressing of magnesium oxide powder prepared by vapour-phase oxidation process

Effect of starting particle size on hot-pressing of magnesium oxide (MgO) powder was examined using seven kinds of MgO powders prepared by a vapour-phase oxidation process; the average primary particle sizes were 11, 25, 32, 44, 57, 107 and 261 nm. These compressed powders (compacts) were hot-pressed at a temperature between 900 and 1300°C. The densifications of these compacts during the hot-pressing proceeded via (i) the sintering of primary particles within secondary particles and the rearrangement of secondary particles/grains (900°C), (ii) the gradual grain growth controlled by the pore migration (900∼1100°C) and (iii) the rapid grain growth due to the active mass transfer (1300°C); the grain sizes of MgO compacts hot-pressed at and below 1100°C were <1 μm, while those at 1300°C attained 20∼30 μm. The transluscent compact with the relative density of 99.7% could be obtained when the compressed powder with the average primary particle size of 44 nm was hot-pressed at a temperature as low as 1100°C for 1 h. This revised version was published online in November 2006 with corrections to the Cover Date.     

Structure and surface properties of diaminocyclohexane plasma polymer films

Plasma polymers of three isomers of diaminocyclohexane (DACH) were deposited on polyethylene, SiO₂, and mica at 20°C. The deposition rate was measured as a function of plasma density and power; a maximum was observed in the latter function. The deposition rate was highest for the monomer with the highest flow rate. The film refractive index was observed to increase with both power density and the degree of fragmentation in the plasma. Film composition was measured by elementary analysis, and was found to be almost identical for each of the three isomers; a mechanism for the polymerization reaction is proposed. The percentage of primary amino groups decreased with increasing power density and with film thickness. Surface force measurements of the thickness and refractive index agreed well with the corresponding ellipsometry values in dry air, and am adhesive force, independent of power density, was measured. When the film was exposed to water vapor, it swelled considerably and the adhesion was determined by capillary forces. Associated with swelling, at high power, was an extremely regular 2-ply rope pattern of protruding material. © 1993 John Wiley & Sons. Inc.     

Practical method for generation expansion planning based on dynamic programming

This paper presents an efficient method which provides the optimal generation mix and the optimal generation construction process. The approximation method in which the dynamic programming technique and gradient method are combined is applied to determine the optimal generation mix with hydropower generation technologies. The successive approximations dynamic programming (SADP) technique, which is very suitable for high-dimensional multistage decision process problems, is used for obtaining the optimal generation construction process. The effectiveness and feasibility of the developed technique are demonstrated on a practical power system model which has five types of generation technologies including a hydropower generation technology.     

Differential activity of the G protein beta5 gamma2 subunit at receptors and effectors

The G protein beta5 subunit differs substantially in amino acid sequence from the other known beta subunits suggesting that beta gamma dimers containing this protein may play specialized roles in cell signaling. To examine the functional properties of the beta5 subunit, recombinant beta5 gamma2 dimers were purified from baculovirus-infected Sf9 insect cells using a strategy based on two affinity tags (hexahistidine and FLAG) engineered into the N terminus of the gamma2 subunit (gamma2HF). The function of the pure beta5 gamma2HF dimers was examined in three assays: activation of pure phospholipase C-beta in lipid vesicles; activation of recombinant, type II adenylyl cyclase expressed in Sf9 cell membranes; and coupling of alpha subunits to the endothelin B (ETB) and M1 muscarinic receptors. In each case, the efficacy of the beta5 gamma2HF dimer was compared with that of the beta1 gamma2HF dimer, which has demonstrated activity in these assays. The beta5 gamma2HF dimer activated phospholipase C-beta with a potency and efficacy similar to that of beta1 gamma2 or beta1 gamma2HF; however, it was markedly less effective than the beta1 gamma2HF or beta1 gamma2 dimer in its ability to activate type II adenylyl cyclase (EC50 of approximately 700 nM versus 25 nM). Both the beta5 gamma2HF and the beta1 gamma2HF dimers supported coupling of M1 muscarinic receptors to the Gq alpha subunit. The ETB receptor coupled effectively to both the Gi and Gq alpha subunits in the presence of the beta1 gamma2HF dimer. In contrast, the beta5 gamma2HF dimer only supported coupling of the Gq alpha subunits to the ETB receptor and did not support coupling of the Gi alpha subunit. These results suggest that the beta5 gamma2HF dimer binds selectively to Gq alpha subunits and does not activate the same set of effectors as dimers containing the beta1 subunit. Overall, the data support a specialized role for the beta5 subunit in cell signaling.     

Protein modification by a Maillard reaction intermediate methylglyoxal. Immunochemical detection of fluorescent 5-methylimidazolone derivatives in vivo

Methylglyoxal (MG), an endogenous metabolite that increases in diabetes, is a common intermediate in nonenzymatic glycation (Maillard reaction) in vivo. Here we describe the immunochemical approach to the detection of MG adducts in proteins in vitro and in atherosclerotic lesions of human aorta in vivo. The reaction of protein (bovine serum albumin) with MG led to selective loss of arginine and lysine residues, accompanied by the formation of 5-methylimidazolone (N delta-(5-methylimidazolon-2-yl)ornithine) and imidazolysine (1,3-di-lysino-4-methylimidazole) derivatives, respectively. The anti-5-methylimidazolone antibody was prepared by immunizing rabbits with a MG-keyhole limpet hemocyanin conjugate and purifying the serum on an affinity gel prepared by covalent attachment of the 5-methylimidazolone derivative. The antibody cross-reacted with the proteins treated with not only MG but trioses, such as hydroxyacetone, dihydroxyacetone, and glyceraldehyde. The immunohistochemical analysis revealed that atherosclerotic lesions of human aorta contained 5-methylimidazolone derivatives whose distributions were identical to those of advanced glycation end products (AGEs) detected by the anti-AGE antibody.     

Growth condition of iodine-doped n+-CdTe layers in metal-organic vapor phase epitaxy

Iodine doping of CdTe layers grown on (100) GaAs by metal-organic vapor phase epitaxy (MOVPE) was studied using diethyltelluride (DETe) and diisopropyltelluride (DiPTe) as tellurium precursors and ethyliodine (EI) as a dopant. Electron densities of doped layers increased gradually with decreasing the growth temperature from 425°C to 325°C. Doped layers grown with DETe had higher electron densities than those grown with DiPTe. When the hot-wall temperature was increased from 200°C to 250°C at the growth temperature of 325°C, doped layers grown with DETe showed an increase of the electron density from 3.7×10¹⁶ cm⁻³ to 2.6×10¹⁸ cm⁻³. On the other hand, such an increase of the electron density was not observed for layers grown with DiPTe. The mechanisms for different doping properties for DETe and DiPTe were studied on the basis of the growth characteristics for these precursors. Higher thermal stability of DETe than that of DiPTe was considered to cause the difference of doping properties. With increasing the hot-wall temperature from 200°C to 250°C, the effective ratio of Cd to Te species on the growth surface became larger for layers grown with DETe than those grown with DiPTe. This was considered to decrease the compensation of doped iodine and to increase the electron density of layers grown with DETe. The effective ratio of Cd to Te species on the growth surface also increased with decreasing growth temperature. This was considered to increase the electron density with decreasing growth temperature.     

Frequency dependence of AC susceptibility of bcc solid3He

We have measured AC susceptibility of bcc solid³ He in a two orders of magnitude with a wider range than in the previous study. For the sample of 23.0 cm³/mol, the relaxation time between the Zeeman and exchange reservoirs in the paramagnetic state is on the order of 10 ms, while the relaxation time in the ordered state decreases down to one hundredth of the value in the paramagnetic state. This small relaxation time is related to the process of spin wave relaxation in the ordered state.