生物物理化学
Online ISSN : 1349-9785
Print ISSN : 0031-9082
ISSN-L : 0031-9082
43 巻, 3 号
選択された号の論文の9件中1~9を表示しています
  • 久保見 朋子, 藤田 剛, 松尾 雄志
    1999 年 43 巻 3 号 p. 129-138
    発行日: 1999/09/15
    公開日: 2009/03/31
    ジャーナル フリー
    FACE (fluorophore-assisted carbohydrate electrophoresis) is a method for the separation of fluorophore-labeled saccharides by the use of polyacrylamide gel electrophoresis. The method is characterized by high resolution, high sensitivity, and ease of use without expensive giant-sized machine such as mass spectrometry. It has been found to be useful for the analysis of oligosaccharides profiling of N-linked or O-linked glycans after releasing saccharides from glycoproteins and for sequencing oligosaccharides component. Judging from the importance of oligosaccharides in glycoproteins and the changes in oligosaccharides of cancer cell surface, the method described here is hopeful as a unique analysis tool in the field of basic medical science.
  • 戸沢 辰雄
    1999 年 43 巻 3 号 p. 139-145
    発行日: 1999/09/15
    公開日: 2009/03/31
    ジャーナル フリー
    It has been proposed that the immunoglobulins in some naturally occurring macroamylases may primarily directed to animal amylase. This would suggest that there may also be an antibody to animal amylase with inability to bind endogenous amylases in circulation. On the basis of this assumption, an antibody to porcine pancreatic amylase (PPA) was investigated in sera from patients with inflammatory bowel disease (IBD): ulcerative colitis (UC) and Crohn's disease (CD), because serum antibodies to various dietary antigens have been demonstrated to be a significantly prevalent in patients with IBD. This investigation took advantage of the abilities of serum to inhibit specific activity of added PPA and to form an artificial macroamylase with the PPA. As expected, such antibodies to PPA were detected frequently in patients with CD (19/50, 38%), rarely in patients with UC (1/50, 2%) and not at all in normal controls (0/30). There were two different types of antibodies differing in their reaction with PPA; one was an enzyme-inhibiting antibody and another an enzyme-noninhibiting antibody. The former seemed to be of monoclonal or oligoclonal IgG whereas the latter was of polyclonal IgG and IgA or polyclonal IgG alone, and both were detected in almost cases. The presence of the antibodies specific for CD suggests that the antibodies may play an etiological role in CD, of which the etiology is still unknown.
  • 草〓 睦子, 藤田 清貴, 櫻林 郁之介, 鈴木 徳和, 寺邑 能実, 岡本 恵智子, 小林 邦彦, 吉岡 尚文, 荒川 正明
    1999 年 43 巻 3 号 p. 147-151
    発行日: 1999/09/15
    公開日: 2009/03/31
    ジャーナル フリー
    Separax-SP膜と反応するIgA1-κ型M-蛋白を見い出し検討を加えた. このM-蛋白は, 2.0M NaClの洗浄でも吸着したセ・ア膜から解離することはなく, 還元アルキル化および neuraminidase 処理後でもセ・ア膜との反応性は消失しなかった. ジアセチルセルロースとトリアセチルセルロースを等量混合し油性成分を含まないセ・ア膜を用い電気泳動を行ったところ, 明瞭なM-蛋白帯は認めず, 原点のα1からβ位に残る幅広い異常蛋白帯が観察され, M-蛋白とアセチルセルロースとの結合による異常が示唆された. 患者IgA1型M-蛋白をプロテアーゼ処理後, 精製したαFab分画, およびαFc分画単独ではセ・ア膜との反応性は消失した. これらの結果から, セ・ア膜との反応は抗原抗体反応によるものではなく, 患者IgA1型M-蛋白がもつ物理的立体構造に起因した結合の可能性が高い.
  • 宮島 絹江, 星野 忠, 吉冨 要子, 阿部 和美, 中村 利弘, 河野 均也, 菰田 二一
    1999 年 43 巻 3 号 p. 153-157
    発行日: 1999/09/15
    公開日: 2009/03/31
    ジャーナル フリー
    われわれは, S1-アミラーゼ (AMY) よりも電気的易動度が遅いS型AMY (slow-S) バリアントを見いだし, 易動度の順に陽極側から slow-S1, slow-S2, slow-S3と命名した. slow-S検体の検出は, 抗ヒトS-AMY阻害モノクローナル抗体処理と未処理試料との電気泳動パターンを比較して, バンドの消失または減少からその同定を行った. 各 slow-Sバンドの易動度は, slow-S1はP2とほぼ同位置に, slow-S2は dominant-P2Sとほぼ同位置に, そして slow-S3はP1と dominant-P2Sの間にそれぞれ検出された. slow-Sの出現頻度は, 7,234検体中, slow-S1は5検体 (0.07%), slow-S2は12検体 (0.17%), slow-S3は2検体 (0.03%) であった. しかしながら, slow-Sバリアント出現の起因については, 現時点では不明であり, 遺伝的な素因, 糖鎖による修飾, デアミノ化の可能性について, 今後検討していく必要がある.
  • 渡部 俊弘, 佐藤 広顕, 相根 義昌, 中澤 洋三, 高野 克己
    1999 年 43 巻 3 号 p. 159-164
    発行日: 1999/09/15
    公開日: 2009/03/31
    ジャーナル フリー
    Phospholipase D (PLD) was highly purified from cabbage leaves of three different cultivars. Each purified PLD showed a single protein band with molecular masses of 87 kDa on SDS-PAGE, respectively. Peptide maps of each purified PLD after digestion with Lysyl Endopeptidase or V8 Protease gave the same pattern of proteolytic peptide fragments. Previously, Pannenberg et al. suggested the existence of two isozymes designated as PLD1 and PLD2. The N-terminal amino acid sequences of whole PLD and of proteolytic peptide fragments were determined. As a result, the PLD isolated from cabbage leaves was identified as PLD2.
  • 渭原 博, 戸谷 夏子, 柿木 孝志, 谷 あすか, 青木 豊, 橋詰 直孝, 吉田 光孝
    1999 年 43 巻 3 号 p. 165-169
    発行日: 1999/09/15
    公開日: 2009/03/31
    ジャーナル フリー
    Electrophoretic separation and quantitation of serum proteins, including paraproteins, remain essential for the diagnosis and management of a variety of disorders. We examined the Beckman automated Paragon 2000 capillary zone electrophoresis (CZE) system and compared its electropherograms to those obtained by cellulose acetate electrophoresis (CAE). Within-run CVs using seven different capillaries and between-day CVs for CZE were <2% for albumin; they were <6% for α1-globulin, α2-globulin, β-globulin, and γ-globulin. From 2.7 to 15.0g/dl of total protein concentrations, the same electrophoretic patterns could be observed. Comparisons of the five principal bands obtained by CZE and cellulose acetate electrophoresis (CAE) gave correlation coefficients from 0.8 to 0.9. On a quantitative basis in 55 specimens, albumin by CZE was on average 10% lower than CAE, and α1-globulin and β-globulin were 93% and 15% higher than by CAE, respectively (p<0.001). Based on our use of the Beckman Paragon software and our reference values (percent of total protein) for CZE of 56.3 to 68.9% for albumin, 3.9 to 6.3% for α1-globulin, 4.8 to 8.4% for α2-globulin, 7.9 to 11.9% for β-globulin, and 9.9 to 21.5% for γ-globulin, we found 75% to 100% agreement with CZE; i.e., both methods gave either normal or abnormal results of the time. Patients showing a gammopathy gave values that were higher by CZE 60% of the time. Furthermore, the CZE system can resolve prealbumin, α1-acid glycoprotein, α1-antitrypsin, hemopexin, transferrin, and complement besides to the usual five bands.
  • 荒木 秀和, 片桐 綾子, 佐野 恵一, 松田 武英, 井村 敏雄, 下条 文武, 片岡 伸久朗
    1999 年 43 巻 3 号 p. 171-175
    発行日: 1999/09/15
    公開日: 2009/03/31
    ジャーナル フリー
    Atypical bands are frequently found at cathode side than the band of apolipoprotein (apo) E4 isoforms separated by Phenotyping Apo E IEF System (Jokoh Co. Ltd.). Then, we developed to resolve the atypical phenotype bands as apo E5 and apo E7 isoforms by PCR with restriction fragment length polymorphism and with amplification refractory mutation system. From the results, the genetic identification of atypical apo E5 and apo E7 phenotype bands was well accorded to the apo E phenotypes separated by the Phenotyping Apo E IEF system.
  • 藤田 清貴, 草〓 睦子, 荒川 正明, 櫻林 郁之介
    1999 年 43 巻 3 号 p. 177-180
    発行日: 1999/09/15
    公開日: 2009/03/31
    ジャーナル フリー
    Immunofixation electrophoresis (IFE) is used routinely to identify monoclonal immunoglobulins (M-proteins) in serum and Bence Jones proteins in urine. A method for subclass typing of IgG and IgA type M-proteins using MINIFIXTM IFE kit is described in this paper. The antisera prepared for MINIFIXTM kit (IgG and IgA subclass sets), were used and M-proteins in 20 patient sera were analyzed. The subclass types of the M-proteins were identified as seven IgG1, one IgG2, two IgG3, seven IgA1, two IgA2 and one IgG1-IgA1 multi type. These results did not conflicted with those of western blotting analysis. The MINIFIXTM IFE kit is useful for identification of the subclasses of IgG and IgA type M-proteins. The MINIFIXTM IFE kit employs regular-sized agarose gels (102×77mm) but because of the reduced track width, will allow two patient samples per gel. Consequently, the MINIFIXTM IFE system offers excellent economy with each kit containing all the necessary reagents.
  • 前田 哲男, 前田 光雄, 戸沢 辰雄
    1999 年 43 巻 3 号 p. 181-183
    発行日: 1999/09/15
    公開日: 2009/03/31
    ジャーナル フリー
    A case of macroamylasemia occurring in a man and his daughter is described. A 65-year-old man who was hospitalized for hepatic dysfunction had persistent hyperamylasemia. The ACCR was low level. Serum amylase electrophoresis showed an abnormal broad band. The amylase elution pattern on a Sephadex G-200 column showed a macromolecular peak between 19S and 7S, indicating that the amylase had a larger molecular structure than that of normal amylase. An immunoprecipitate examination revealed the macroamylase to be an amylase linked to immunoglobulin A-λ. Serum amylase from the patient's healthy daughter showed the same characteristics as the patient, although amylase from the other members of his family did not. The etiology of naturally occurring macroamylasemia has not been resolved.
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