生物物理化学
Online ISSN : 1349-9785
Print ISSN : 0031-9082
ISSN-L : 0031-9082
51 巻, 2 号
選択された号の論文の11件中1~11を表示しています
総説
  • 田中 啓二
    原稿種別: 特別講演
    2007 年 51 巻 2 号 p. 83-89
    発行日: 2007年
    公開日: 2009/12/04
    ジャーナル フリー
    Ubiquitin, an 8.6-kDa highly conserved protein, is a landmark molecule functioning as a posttranslational modifier. It is covalently attached to target proteins in a fashion of polymerization, resulting in a poly-ubiquitin chain that serves as a degradation signal. The 26S proteasome is an eukaryotic ATP-dependent protease responsible for selective degradation of the polyubiquitin-tagged proteins in eukaryotic cells. It appears to act as an elegantly organized apparatus designed for efficient and exhaustive hydrolysis of proteins, and can in fact be regarded as a protein-destroying machinery. To date, there are growing lines of evidence addressing the importance of the ubiquitin-proteasome system that catalyzes various biological reactions rapidly, orderly, exhaustively, and unidirectionally. On the other hand, autophagy (Greek for “self-eating”) is an evolutionarily conserved pathway in which the cytoplasm and organelles are engulfed within double-membraned vesicles, known as autophagosomes, which rapidly fuse with lysosomes and their contents together with the inner membrane are degraded by a variety of lysosomal digestive enzymes. While autophagy has been thought to contribute to bulk degradation non-selectively, but recent genetic analaysis with mice reveals that ablation of autophagy leads to accumulation of ubiquitin-positive inclusions, implying that the novel role of ubiquitin is to provide a signal that shuttles ubiquitinated proteins for autophagy. Currently, the roles of ubiqutin are expanding in various fields of life science. In this review, I will review the ubiquitin-mediated proteolysis pathway with a special reference to proteasomes and autophagy, focusing on how ubiquitin is linked to the pathogenesis of neurodegenerative diseases.
  • 菅野 剛史, 青木 芳和
    原稿種別: 〔教育講演〕
    2007 年 51 巻 2 号 p. 91-97
    発行日: 2007年
    公開日: 2009/12/04
    ジャーナル フリー
    The main separation procedures used by clinical laboratories to assist in diagnosis are summarized. Separation of serum proteins and isozymes by electrophoresis and separation of hemoglobin A1c and plasma lipoproteins by high performance liquid chromatography (HPLC) are discussed as examples. An automated system for serum protein electrophoresis was developed. We, then, designed an overlapping display system that monitors change in protein electrophoresis. With respect to diagnostic use of isozymes, isozyme analysis proved to be the key to the first diagnosis of lactate dehydrogenase-M subunit deficiency. Since then, more than 30 cases involving different mutations have been found worldwide. With respect to HPLC, specific column preparations have made it possible to quickly separate hemoglobin A1c from the hemoglobin fraction. By this separation, many hereditary hemoglobin abnormalities can be detected. Hemoglobin Sagami was the first hemoglobin subtype that could not be identified by the commonly used propan2ol test and isoelectrofocucing. Moreover, HPLC is a powerful tool for separation of lipoprotein subclasses and can be used to detect atherosclerotic stage in patients with ischemia. In many clinical cases, use of one of these separation procedures is essential for the determination of pathophysiologic stage.
ワークショップ:タンパク分画用の新規セルロースアセテート膜,セレカVSPの性能評価
  • 前川 真人
    原稿種別: 第57回日本電気泳動学会総会ワークショップ
    2007 年 51 巻 2 号 p. 99
    発行日: 2007年
    公開日: 2009/12/04
    ジャーナル フリー
  • 岡田 英孝, 新城 俊憲
    原稿種別: 〔ワークショップ:タンパク分画用の新規セルロースアセテート膜,セレカVSPの性能評価〕
    2007 年 51 巻 2 号 p. 101-103
    発行日: 2007年
    公開日: 2009/12/04
    ジャーナル フリー
    The stable supply of the cellulose acetate membrane (SEPARAX-SP) for electrophoresis support media became difficult with production device deterioration. Therefore, Toyo Roshi Kaisha, Ltd. was transferred production technology from Fuji Co. and tried to develop a new membrane with similar performance. At the beginning physical properties of the membrane differed slightly possibly due to the difference of production device; however, recently a new membrane with similar performance to SEPARAX-SP has been completed and named SELECA-VSP.
    Physical properties, such as tearing strength and elasticity, electrophoretic properties of SELECA-VSP were similar to those of SEPARAX-SP. By scanning electron microscope, SELECA-VSP membrane surface and cross section showed similar image and uniformly pored.
    To investigate a practical use of SELECA-VSP in clinical laboratory, a working group was organized by Japanese Electrophoresis Society and has examined basic and clinical performance of SELECA-VSP. Because the results have been revealed almost good performance, we go on developing for placing SELECA-VSP on the market.
  • 小野寺 和彦, 石原 清隆, 有園 秀敏
    原稿種別: 〔ワークショップ:タンパク分画用の新規セルロースアセテート膜,セレカVSPの性能評価〕
    2007 年 51 巻 2 号 p. 105-107
    発行日: 2007年
    公開日: 2009/12/04
    ジャーナル フリー
    As a cellulose acetate membrane for serum protein fraction test, SELECA-VSP has been developed to substitute for SEPARAX-SP. Then we investigated whether SELECA-VSP can be used for protein fraction test by use of full-automated electrophoresis system (JOKOH CTE series).
    A difference was observed in the A / D sensor level detecting that SELECA-VSP fed into buffer bath because SELECA-VSP was thinner than SEPARAX-SP. Therefore, the adjustment of the A / D sensor level is possibly required when SELECA-VSP is substituted for SEPARAX-SP.
    On the other hand, there were no difference in membrane feeding test, electrophoretic properties and measured percentage of each protein fraction of control serum. Slight change in the full-automated electrophoresis system parameter easily leads to apply SELECA-VSP to routine laboratory test using JOKOH CTE series electrophoresis system.
  • 古澤 幸弘, 三浦 弘一
    原稿種別: 〔ワークショップ:タンパク分画用の新規セルロースアセテート膜,セレカVSPの性能評価〕
    2007 年 51 巻 2 号 p. 109-112
    発行日: 2007年
    公開日: 2009/12/04
    ジャーナル フリー
    As a cellulose acetate membrane for serum protein fractionation testing, SELECA-VSP has been developed to substitute for SEPARAX-SP. In order to investigate whether or not SELECA-VSP is available for routine laboratory testing, we evaluated basic performance of SELECA-VSP by use of automated electrophoresis system, Olympus AES series.
    As for the consecutive transportability, no jam was occurred on 300 consecutive membrane sheets of SELECA-VSP, lot T13-2. Reproducibility was good in within-run and between-run precision. However, the waiting coefficient for application should be set up adequately in each laboratory because SELECA-VSP tends not to dry up. Using SELECA-VSP the decolored clear zone between β globulin fraction and γ globulin fraction is more distinct than SEPARAX-SP, the results analyzed by diagnosis support program may be misinterpreted. In order to avoid the misinterpretation, the standard densitogram of normal control by use of SELECA-VSP should be prepared.
  • 宮﨑 京子, 米山 正芳, 高橋 美穂, 江上 照夫, 大西 宏明, 渡邊 卓
    原稿種別: 〔ワークショップ:タンパク分画用の新規セルロースアセテート膜,セレカVSPの性能評価〕
    2007 年 51 巻 2 号 p. 113-117
    発行日: 2007年
    公開日: 2009/12/04
    ジャーナル フリー
    Recently a new cellulose acetate membrane for electrophoresis to analyze protein fractions, named SELECA-VSP, was developed by Toyo Roshi Kaisha Ltd. In advance of commercial supply, a study group was organized to investigate utility of this new membrane in comparison with SEPARAX-SP, a standard membrane for protein fraction assay widely used in Japan. As a member of this study group, our laboratory mainly investigated the difference between the two membranes when analyzing protein fractions in serum as well as ascites, pleural fluid, cerebrospinal fluid and urine. JOKO-CTE8000 was used as a protein fraction analyzer. When buffer lacking EDTA was used in electrophoresis of serum and ascites, the correlation efficiency between the two membranes in percentage of the β fraction was inferior to that of other fractions. When EDTA-containing buffer was used in analysis of serum and ascites, however, the correlation between the two membranes in β-fraction was improved to a comparable level to other fractions. The correlations between the two membranes were satisfactory in analysis of samples other than serum and ascites, regardless of the presence of EDTA in buffer. We conclude that this newly developed membrane has the similar performance to a standard membrane and could be applied in assay of protein fraction in clinical samples.
  • 中村 和代, 須郷 秋恵, 宮島 栄治
    原稿種別: 〔ワークショップ:タンパク分画用の新規セルロースアセテート膜,セレカVSPの性能評価〕
    2007 年 51 巻 2 号 p. 119-123
    発行日: 2007年
    公開日: 2009/12/04
    ジャーナル フリー
    Following the discontinuation of manufacturing SEPARAX-SP, a cellulose acetate membrane from Fujifilm which had long been used in clinical laboratory testing, Advantec Toyo decided to manufacture another cellulose acetate membrane that exhibits the properties of SEPARAX-SP. However, it is unclear whether SELECA-VSP, the new product based on certain changes in the manufacturing process of the cellulose acetate membrane, is equivalent to SEPARAX-SP in quality. Thus, we conducted a basic study using a prototype membrane of SELECA-VSP on an automatic electrophoresis apparatus, and found that it could be applied to the currently used model of automatic electrophoresis apparatus without adjusting operating conditions. The basic study results and their correlations were promising enough to suggest that SELECA-VSP could replace SEPARAX-SP without problems.
  • 渡邊 弘子, 杉浦 綾, 前川 真人
    原稿種別: 〔ワークショップ:タンパク分画用の新規セルロースアセテート膜,セレカVSPの性能評価〕
    2007 年 51 巻 2 号 p. 125-128
    発行日: 2007年
    公開日: 2009/12/04
    ジャーナル フリー
    Serum protein fractionation has been performed by electrophoresis on cellulose acetate membrane in clinical laboratory and has been applied to diagnostic procedure for pathological states. In Japan SEPARAX-SP (Fuji film Co.) has been used as the supporting media of cellulose acetate membrane for about twenty years and will finish its production soon. A new cellulose acetate membrane, SELECA-VSP (Toyo Roshi Kaisha Ltd.) has been developed instead of SEPARAX-SP. The SELECA-VSP has almost similar features and function. In this study we evaluated quality and utility of SELECA-VSP by automated electrophoresis system (Olympus AES320). Protein fractionation using SELECA-VSP was not worse than that of SEPARAX-SP. Separation sensitivity was higher in SELECA-VSP. However, irregularly smeared bands were sometimes observed adjacent to monoclonal protein bands. Areas of each fraction are highly correlated in between SEPARAX-SP and SELECA-VSP. The fraction area of α1-globulin was significantly different in SEPARAX-SP and SELECA-VSP, and about 0.01 g / dl higher in SELECA-VSP. After the presence of the difference is recognized, automated classification system for pathological states by protein fractionation could be utilized for routine laboratory testing.
    In conclusion, the new cellulose acetate membrane, SELECA-VSP will replace SEPARAX-SP as electrophoretic supporting media for serum protein fractionation.
  • 大竹 和子, 大竹 皓子, 深田 比呂子, 堀井 康司, 村田 満
    原稿種別: 〔ワークショップ:タンパク分画用の新規セルロースアセテート膜,セレカVSPの性能評価〕
    2007 年 51 巻 2 号 p. 129-133
    発行日: 2007年
    公開日: 2009/12/04
    ジャーナル フリー
    New cellulose acetate membrane SELECA-VSP was prepared as a substitute for the conventional cellulose acetate membrane SEPARAX-SP, which has been widely used for serum protein electrophoretic fractionation. We evaluated the separating quality of this new membrane, using fully automated serum protein electrophoresis system AES630.
    Correlation of serum protein fraction values (percentage composition) between the two membranes, those patterns of sera from patients with protein disorders, and reference intervals in healthy adults were analyzed. In addition, the electrophoretic determinations of amylases and glycoproteins were tested.
    The correlation coefficients of the each electrophoretic fraction between SELECA-VSP and of SEPARAX-SP were greater than 0.97. Reference intervals in healthy adults on SELECA-VSP and on SEPARAX-SP were nearly equal.
    The separating performance of SELECA-VSP were rather higher as compared to SEPARAX-SP for serum protein electrophoretic fractionation, amylase isoenzymes and glycoproteins.
    The utility and effectiveness of a new membrane SELECA-VSP can be best appreciated on the basis of our experimental results.
  • 渡邊 弘子, 杉浦 綾, 大竹 和子, 大竹 皓子, 深田 比呂子, 堀井 康司, 村田 満, 中村 和代, 須郷 秋恵, 宮島 栄治, 宮 ...
    原稿種別: 〔ワークショップ:タンパク分画用の新規セルロースアセテート膜,セレカVSPの性能評価〕
    2007 年 51 巻 2 号 p. 135-145
    発行日: 2007年
    公開日: 2009/12/04
    ジャーナル フリー
    In order to substitute SELECA-VSP for SEPARAX-SP, we investigated membrane performance difference among SELECA-VSP lots and among laboratories. We examined three lots of SELECA-VSP, namely T10, T12 and T14. Automated electrophoresis system for protein fraction test used are Olympus AES series in two laboratories and JOKOH CTE series in other two laboratories. The 4 laboratories assayed common serum samples as usual procedure of each laboratory.
    Among different membrane lots, good correlation between each two lot was observed in the five serum protein fractions. There was statistically significant difference among the membrane lots, however, it might be only due to good reproducibility and statistics. The bias was below clinical permission limit.
    Among laboratories, good correlation between each two laboratory was observed in the five serum protein fractions. The bias between laboratories is below permission limit, compared with bias detected from external quality assessment program for SEPARAX-SP.
    In conclusion, because the bias in membrane lots and laboratories can be ignored for laboratory diagnosis of serum protein fraction test, SELECA-VSP will come onto the market to make up for SEPARAX-SP.
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