Accumulation of Misfolded Protein Aggregates Leads to the Formation of Russell Body-like Dilated Endoplasmic Reticulum in Yeast

RNAP-I, an aspartic proteinase from a filamentous fungus Rhizopus niveus, is secreted very efficiently in Saccharomyces cerevisiae. It is synthesized first as a precursor form with signal sequence and prosequence in its amino-terminus. Our previous study indicated that the prosequence of RNAP-I had important roles in its correct folding and secretion in yeast, and that a prosequence-deleted derivative of RNAP-I, Δpro, was not secreted but was retained and degraded in the yeast endoplasmic reticulum (ER). In the present study, we show that the accumulation of Δpro in the yeast ER caused elevated synthesis of ER resident chaperones, indicating that Δpro is recognized as an unfolded protein species in the ER. Our biochemical data demonstrated that Δpro formed aggregates which contained BiP, but not protein disulfide isomerase (PDI), in the ER. Immunoelectron microscopical analysis revealed that the Δpro aggregates were indeed visible as electron-dense regions in the ER and nuclear envelope. Such ‘chaperone-associated misfolded protein bodies’ were observed for the first time in yeast. Morphologies of the ER and nucleus were drastically altered by the accumulation of the Δpro aggregates. The ER lost its flat cisternal shape; the ER lumen extended aberrantly and the ER membrane irregularly proliferated. The misfolded Δpro proteins are probably sorted from the ordinary ER lumen to form the aggregates so that the ER function would not be grossly impaired, and the dilated ER may represent an ER subcompartment where the Δpro aggregates are degraded. © 1997 by John Wiley & Sons, Ltd.     

Surface modification of poly(aryl ether ether ketone) film by remote oxygen plasma

Surface modification of poly(aryl ether ether ketone) (PEEK) film surfaces by oxygen plasma treatment was investigated. Two procedures, the direct plasma treatment and the remote oxygen plasma treatment, were used as oxygen plasma treatments, and the efficiency of the hydrophilic modification was discussed. The direct and remote oxygen plasma treatments lead to degradation of the PEEK film as well as hydrophilic surface modification. The degradation disturbs the surface modification. The remote oxygen plasma treatment rather than the direct oxygen plasma is suitable for the hydrophilic surface modification of the PEEK film. The remote oxygen plasma treatment at 10 W for 60 s forms predominantly C—O groups rather than C=O groups as an oxygen-containing group on the PEEK surface and gives a highly hydrophilic surface with a contact angle of 44 degrees. © 1998 John Wiley & Sons, Inc. J Appl Polm Sci 68:271–279, 1998     

Recovery of serum proteins using cellulosic affinity membrane modified by immobilization of CU2+ ion

An affinity membrane was prepared from a porous cellulose membrane, and adsorption and recovery of serum proteins were investigated from the viewpoint that affinity membranes are efficacious against separation and purification of biomaterials. Into the cellulose membrane, iminodiacetate (IDA) group that acts as a ligand to metal ions was introduced (Cell–IDA membrane), and then Cu²⁺ ion was immobilized (Cell–IDA–Cu membrane). Bovine serum albumin (BSA) and γ-globulin (BγG), which are the major proteins in blood, were adopted as model proteins to be separated. The Cell–IDA–Cu membrane had large adsorption capacity for these proteins despite the low degree of modification. The amounts of proteins adsorbed on the Cell–IDA–Cu membrane increased with increasing pH, and BγG was adsorbed more than BSA. High protein recoveries from the Cell–IDA–Cu membrane were obtained. The separation of these proteins was also conducted under the optimum conditions of adsorption and recovery, and BγG was concentrated more than BSA although the initial concentration of BγG was lower than that of BSA. © 1996 John Wiley & Sons, Inc.     

Detection of pathogenic bacteria by using zinc finger protein fused with firefly luciferase

We constructed a novel bacterial genome detection system using zinc finger protein (ZF) fused with firefly luciferase (ZF-luciferase). Taking advantage of the direct recognition of double-stranded DNA (dsDNA) by ZF, we previously constructed bacterial genome detection systems that did not require dehybridization processes. To detect polymerase chain reaction (PCR) products rapidly and with a high sensitivity, we constructed two kinds of ZF-luciferase, Sp1-fused luciferase (Sp1-luciferase), and Zif268-fused luciferase (Zif268-luciferase). ZF-luciferase not only maintains luciferase activity but also shows dsDNA-binding ability and specificity. Furthermore, we succeeded in the detection of 10 copies of the genome of Legionella pneumophila and Escherichia coli O157. ZF-luciferase would be a useful tool for highly sensitive detection of pathogenic bacterial genome.     

Evolution of long-period stacking ordered structures on annealing as-cast Mg85Y9Zn6 alloy ingot observed by synchrotron radiation small-angle scattering

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Nickel/Zinc Chloride‐Promoted Domino Reaction of Enynes and Enones Including Unstrained C?C Bond Cleavage

Unstrained C C bond cleavage proceeds during the domino reaction of enynes and enones that includes successive C C bond formation under the nickel/zinc/zinc chloride system. The cleavage occurs through β‐syn‐elimination of the 1,3‐dicarbonyl part. In addition, β‐carbon elimination is selective, unlike the β‐hydrogen elimination in the presence of excess zinc chloride.     

Selective production of lactic acid in continuous anaerobic acidogenesis by extremely low pH operation

The selective production of lactic acid by anaerobic acidogenesis with low pH control was examined using a chemostat culture. By decreasing culture pH to 3.5 in a chemostat culture containing mixed microbial populations for anaerobic acidogenesis, heterolactic fermentation became dominant, resulting in the selective production of lactic acid and ethanol. This phenomenon was reversible between the acidic and neutral conditions, and was not affected by the dilution rate. The extremely low pH operation was effective for selective lactic acid production in anaerobic acidogenesis.     

Temperature dependence and fracture criterion of mixed mode I/II fracture toughness of phenolic resin for friction material

In this study, the temperature dependence of the mixed‐mode fracture toughness of the phenolic resin for friction materials is investigated. For pure mode I, the fracture toughness decreases as the temperature increases, and it increases again after showing its minimum value. For pure mode II, the fracture toughness shows a similar trend but has its minimum value at a higher temperature. The temperature dependence of the mixed‐mode fracture toughness varies depending on the mode mixity, which is attributed to the different sensitivity to the relaxation phenomenon. At room temperature, as the fracture toughness for pure mode I and II are almost the same, the fracture locus shows a circular arc. At elevated temperatures, the locus becomes smaller and noncircular. At high temperature, the fracture locus shows an elliptical arc, where the fracture toughness for pure mode II is smaller than that for mode I. An empirical fracture criterion based on the time‐temperature dependence of the resin is proposed, and the proposed method successfully predicts the fracture toughness under various conditions of the temperature, time, and mode mixity. The crack initiation angles, on the other hand, are almost consistent regardless of the temperature, which agree with the maximum hoop stress theory. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011