生物物理化学
Online ISSN : 1349-9785
Print ISSN : 0031-9082
ISSN-L : 0031-9082
43 巻, 2 号
選択された号の論文の13件中1~13を表示しています
  • 斎藤 春雄, 武川 睦寛
    1999 年 43 巻 2 号 p. 49-55
    発行日: 1999/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    ストレス応答MAPK (p38およびJNK) 情報伝達系は種々の物理化学的ストレスにより活性化され, ストレスを被った細胞にアポトーシスを誘導する. この情報伝達系の制御機構を明らかにするため, ヒト経路に相補的な出芽酵母のHOG1カスケードの変異株を用いて, ヒト遺伝子の機能的スクリーニングを行った. その結果, 酵母のMAPKKKに高い相同性を有し, p38およびJNKカスケードを特異的に活性化するヒトMAPKKK, MTK1を得た. また, ストレス経路の抑制分子としてPP2Cαを同定した. さらにMTK1の活性化機構を明らかにするために,two-hybrid 法を用いて, MTK1の制御領域に結合する分子のクローニングを行った. この結果, GADD45と高い相同性をもつ2種の新しい分子を得た. これら3つのGADD45関連遺伝子はストレス刺激により発現が誘導され, MTK1の活性化を介してストレス応答MAPK情報伝達系を活性化し, 細胞にアポトーシスを誘導すると考えられる.
  • 大木 操
    1999 年 43 巻 2 号 p. 57-61
    発行日: 1999/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    We have isolated the NotI sites that exist on the long arms of human chromosomes 21 and 11 as NotI linking clones and constructed complete NotI restriction maps spanning the entire regions. These maps, which provide the most reliable ordering and distance estimation in the regions from pericentromeric loci to the termini, have led to the identification of four fusion genes including AML1-MTG8 in the breakpoints of chromosome translocations associated with myeloid leukemia.
  • マトリライシン, トリプシン, およびラミニン5の機能
    宮崎 香, 宮田 智, 越川 直彦, 水島 寛人, 安光 英太郎
    1999 年 43 巻 2 号 p. 63-67
    発行日: 1999/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    During complex process of tumor metastasis, tumor cells interact with various extracellular matrix proteins. These proteins regulate tumor invasion and metastasis positively or negatively. Tumor cells utilize various types of matrix-degrading proteinases to invade through basement membranes and connective tissues. These matrix proteinases are classified into two major groups: the matrix metalloproteinases (MMPs) such as gelatinases A and B, interstitial collagenase, stromelysin, matrilysin and MT-MMPs, and the matrix serine proteinases such as plasminogen activators, plasmin and trypsin. On the other hand, we have found laminin-5 (ladsin) as a tumor-derived cell scattering factor with potent cell adhesion and cell motility activities. This protein is assumed to contribute to tumor cell migration. This article describes our recent results on functions of matrilysin, trypsin and laminin-5 in the malignant growth of tumor cells.
  • 櫻林 郁之介
    1999 年 43 巻 2 号 p. 69-73
    発行日: 1999/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    Serum protein fractionation using capillary zone electrophoresis and the application is described. There is 20-30 major components in human serum protein, even though the several hundred proteins are exist. However, in serum protein fractionation by cellulose acetate membrane electrophoresis applied to the clinically, it is only five fractions which intermingle many proteins ingredients with one fraction. Clinically, combining change of the 5 fractions has been made to be the assistance of the diagnosis. In the meantime, capillary zone electrophoresis for excellent separation can be consequentially divided the serum proteins into 10 fractions. However, it becomes a result of being inconvenient on the contrary. The clinical significance equal to conventional electrophoresis of protein would be found when divided protein fractions are specially integrated for 5 fractions. The purpose was achieved by the CZE 2000 instrument (Beckman-Coulter Co., USA) and was obtained by the preparation of the software which arranges the serum protein fractions are originally divided into 10 peaks for 5 fractionation. Therefore, it is possible that can fundamentally follow clinical significance by cellulose acetate membrane electrophoresis and moreover, the data of 10 fractionation is also taken out. And, for the identification of regarded M-protein in cases of multiple myeloma and MGUS (monoclonal gammopathy undetermined significance), it is conveniently possible by subtraction method using the capillary electrophoresis. There is dissociation of α1 fraction value between cellulose acetate membrane electrophoresis and capillary electrophoresis. From our detailed experimental results, it is proved that is due to destaining of α1-acid glycoprotein.
  • 大塚 浩二, 寺部 茂
    1999 年 43 巻 2 号 p. 75-78
    発行日: 1999/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    A brief review of interfaces for on-line coupling of capillary electrophoresis (CE) and mass spectrometry (MS) is presented. To enable the CE-MS system, electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) were introduced as MS interfaces for connecting with CE instruments, while the partial-filling method was developed to suppress the introduction of non-volatile components or additives in CE buffers as well as ionic species such as ionic micelles into the MS interface. Strategy for the coupling of micellar electrokinetic chromatography (MEKC) and MS is also discussed. Examples of CE-MS are briefly shown.
  • 中西 豊文
    1999 年 43 巻 2 号 p. 79-84
    発行日: 1999/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    Progress in medicine and molecular research has been increased by the need for simple and reliable methods of detection and characterization of aberrant proteins caused either by mutation or by post-translational modification. We applied electrospray ionization mass spectrometry (ESIMS) to detect and characterize abnormal structure proteins, and to determine the ratio of mutant and wild type proteins. Direct examination of hemolysate might well lead to rapid ascertainment of variant hemoglobins (Hbs) provided that the mass difference between the normal and the abnormal chain is larger than the resolution power of standard instruments. Various procedures are required in the presentation of most plasma and cell proteins other than Hb prior to MS. Two simple methods were deviced to prepare proteins. One was immunoprecipitation with antisera against target proteins, followed by reversed-phase HPLC/ESIMS, and the other was 2-dimensional LC (a strong anion exchange region and a reversed-phase resin) connected to ESIMS. With both methods, the ions of intact normal and variant proteins were clearly observed in samples from patients with neurodegenerative diseases such as familial amyloidotic polyneuropathy (FAP) and amyotrophic lateral screlosis (FALS). Using these procedures, we detected more than 40 cases of transthyretins (TTRs) mutants including to 10 different types. Two of these, [101 Gly->Ser] and [38 Asp->Ala], were new. We also successfully detected 4 different mutants of Cu/Zn-binding superoxide dismutase inerythrocytes, and a variant in spinal cord from patients with FALS by immunoprecipitation mehods. Two of these, [4Ala->Ser] and [111Cys->Tyr], were new. Finally we also measured glycated β-globin N-ternimus hexapeptide (HbA1c) by Poroszyme V8 protease digestion and nanoESIMS technique.
  • 馬場 嘉信
    1999 年 43 巻 2 号 p. 85-89
    発行日: 1999/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    コンピュータにおける半導体集積技術によって培われた, 微細加工技術をバイオテクノロジーに応用したマイクロチップ技術について解説した. まず, バイオテクノロジーに応用できる微細加工技術として, 光リソグラフィー・化学エッチングとLIGAプロセスについて解説し, これらの技術で作製されたマイクロチャンネルとそれを利用したキャピラリー電気泳動によるDNA解析の高速化について紹介した. また, マイクロチップ上でのPCRとキャピラリー電気泳動の統合化とマイクロチップ上のキャピラリー電気泳動と質量分析計の結合によるゲノム解析・プロテオーム解析への応用について概説した.
  • 稲澤 譲治
    1999 年 43 巻 2 号 p. 91-95
    発行日: 1999/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    Comparative genomic hybridization (CGH) is a powerful technique to identify novel amplification sites through all human chromosomes. We employed CGH to explore genomic imbalances in 19 malignant fibrous histiocytomas (MFHs). Together with losses and gains in various chromosome regions, distinct high-level amplifications were found in six loci (4q12-21, 8p21-pter, 8q24.1-qter, 9p12-13, 12p11.2-pter, and 15q11.2-15), suggesting that those regions may contain unknown genes responsible for carcinogenesis. We focused on the 8p amplicon, and a novel gene designated MASL1 (MFH-amplified sequences with leucine-rich repeats 1) was isolated from within this narrowly defined region. Expression of the MASL1 gene was enchanced significantly in MFH tumors bearing the 8p amplicon. The primary structure of its deduced product revealed an ATP/GTP-binding site, three leucine zipper domains, and a leucine-rich tandem repeat, all of which are important structural or functional elements for interactions among proteins related to the cell cycle. These features suggest that overexpression of MASL1 might well be oncogenic with respect to MFH.
  • 石川 冬木
    1999 年 43 巻 2 号 p. 97-102
    発行日: 1999/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    The chromosome ends, called telomeres, are essential for stable transmission of chromosomes. Recent progress in molecular understanding of telomere structures and functions have enabled us to reason how telomere insufficiencies lead to chromosome instability. Examples that can be explained by this hypothesis include cellular aging and cancer progression.
  • 守内 哲也, 古内 恵司, 八島 秀則, 青山 哲也, 外木 秀文, 浜田 淳一
    1999 年 43 巻 2 号 p. 103-106
    発行日: 1999/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    Recent research on genetic alterations found in human cancers has shown that inactivation of a tumor suppressor gene is a major step in the development of cancers. Two strategies have so far been widely used to study tumor suppressor genes in clinical materials: screening with antibodies and DNA structure-based screening. Recently, a different approach based on a biological assay for tumor suppressor gene function has been devised by using yeast as a living test tube. This method takes advantage of the fact that gapped plasmid is repaired by homologous recombination between the linear plasmid and DNA fragment in yeast. The yeast assay in which yeast change color according to the status of tumor suppressor gene raises the very real possibility of simple and large-scale screening for the genetic mutations in clinical samples. We describe here two conceptually different yeast assays: p53 yeast functional assay and APC premature termination assay.
  • 伊東 文生, 山本 博幸, 日野田 裕治, 今井 浩三
    1999 年 43 巻 2 号 p. 107-109
    発行日: 1999/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    Widespread microsatellite instability (MSI) due to the defective DNA mismatch repair underlies the pathogenesis of the majority of hereditary non-polyposis colorectal cancer and a subset of various sporadic malignant tumors. Using 5 microsatellite markers and the criteria of MSI proposed by the National Cancer Institute (NCI) workshop, we analyzed 205 gastric adenocarcinomas for MSI. Based on the number of markers showing instability per tumor, the tumors were divided into three groups; those with two or more of the five markers displaying instability (high MSI, MSI-H), those with one of five markers displaying instability (low MSI, MSI-L), and those with no instability (microsatellite stable, MSS). Among 205 tumors, 30 (15%) were MSI-H. 15 (7%) were MSI-L, and 160 (78%) were MSS. All of the 30 MSI-H tumors demonstrated instability at BAT26, a sensitive marker for the widespread MSI, while none of the 15 MSI-L tumors did. MSI-H tumors were significantly associated with distal location and well or moderate differentiation, but MSI-H tumors were indistinguishable from MSS tumors. Bax frameshift mutations were detected in 60% of the 30 MSI-H tumors, while not in any of the 15 MSI-L tumors. These results suggest that microsatellite analysis using the criteria proposed by the NCI workshop may appropriate for gastric cancers because it unveils real differences in genotype and phenotype.
  • 岡崎 登志夫, 中川 孝幸, 鎌倉 智行, 柳澤 裕子, 長井 辰男, 菅野 剛史
    1999 年 43 巻 2 号 p. 111-115
    発行日: 1999/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    血清中で, 遊離Hbから速やかにヘムが解離し, アルブミンなどと結合した. しかし, Hp-Hb複合体は, ヘムを放さなかった. ザイモグラフィーや脂肪酸添加試験によって, 血清中におけるHbからのヘム解離は, 脂肪酸によって促進されることが証明された.
  • 川野 克己, 日比 望, 久保野 勝男, 谷亀 光則, 堺 秀人, 石井 直明, 塚田 裕
    1999 年 43 巻 2 号 p. 117-121
    発行日: 1999/06/15
    公開日: 2009/03/31
    ジャーナル フリー
    Advanced Glycation End products (AGEs) は, メイラード反応後期生成物とも呼ばれ, 糖尿病性合併症の原因物質の一つではないかと考えられている. われわれは, 血清中AGE測定系を確立し, 糖尿病さらにその合併症進展と血中AGE量との関係等について検討を行ってきた. 今回, 腹膜透析液を用い, ゲル濾過, affinity chromatography, western blotting 等の手法により, AGEの血中存在様式について検討を行った. その結果, AGE構造の大部分は, 血清蛋白と結合していることが明らかとなった. さらに, AGE化されている蛋白成分としてIgG, アルブミン, β2-マイクログロブリンを同定することができた.
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