Clinical Application of Human alpha_1-T Glycoprotein

人α_1-T糖蛋白的临床应用

• 批准号: 62571037
• 负责人: ITOH Yoshihisa
• 金额: $ 0.83万
• 依托单位: Jichi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62571037/
• 关键词: alpha_1-T glycoprotein; Column Chromatographies; Ammonium sulfate Precipitation; Chronic Renal Failure (CRF); Hepatic Dysfunction; EIA; 単純免疫拡散法; α_1ーT glycoprotein; ユリンエステラーゼ; α^1-Tグリコプロティン; 肝疾患

Human alpha_1-T glycoprotein (alpha_1-T) was a glycoprotein initially isolated from serum by Heide and Haupt in 1964 by rivanol-ammonium sulfate precipitation. The present author successfully purified this protein from serum with the use of modern chromatographic techniques and determined some physicochemical properties as well as partial amino acid sequence. Using established assays of single radial immunodiffusion and qnzyme immunoassay, clinical significance of this protein was investigated. The outline of the present study was summarized as follows.I. Establishment of alpha_1-T Purification: Pooled normal sera were precipitated with 45% ammonium sulfate and its supernatent was further saturated to 75%, precipitate of which was used as a starting material. In an initial purification alpha_1-T was isolated by 5 steps of column chromatographies including ConA affinity, anti-albumin attached affinity, Blue Sepharose, gel-filtration on Sephadex G-200, and cation exchange on CM Sephadex … More C-50. After the preparation of mouse monoclonal antibody these 5 steps were reduced to 3 steps while keeping high yield and purity. Purified alpha_1-T was demonstrated to be a molecular weight of 77,500 daltons, PI 4.5, and tryptophan poor content in amino acid composition. A partial amino acid sequence was determined which showed no any homology exsisting proteins so far reported.II. Clinical Application: The serum level in normal individuals aged from 10 to 60 years old was 71.8 12.3 mg/l without age and sex differences, while that in various diseases was low in cases with hepatic dysfunction and CRF. Of note worth is that the value was rather elevated in acute hepatitis with convalescent stage and drug-induced hepatitis with recovery. These results indicated that this protein is mainly produced in the liver and the measurement of this protein is useful for the evaluation of parenchymal liver cell damage and its repair. The study on low molecular proteins was also facilitated by the present project. Less

人alpha_1-T糖蛋白（Human alpha_1-T glycoprotein，简称alpha_1-T）是Heide和Haupt于1964年首次用利凡诺-硫酸铵沉淀法从血清中分离得到的一种糖蛋白。本文利用现代色谱技术成功地从血清中纯化了该蛋白，并测定了其理化性质和部分氨基酸序列。采用已建立的单径向免疫扩散法和酶免疫分析法，探讨该蛋白的临床意义。本研究的大纲总结如下：α _1- t纯化的建立：将正常血清池用45%硫酸铵沉淀，上清进一步饱和至75%，沉淀作为起始原料。在初始纯化过程中，通过ConA亲和层析、抗白蛋白亲和层析、Blue Sepharose层析、Sephadex G-200层析和CM Sephadex…More C-50层析进行分离。制备小鼠单克隆抗体后，在保持高产率和高纯度的前提下，将这5步减少为3步。纯化后的alpha_1-T分子量为77500道尔顿，PI为4.5，氨基酸组成中色氨酸含量较低。测定了部分氨基酸序列，发现与已有报道的蛋白质无同源性。临床应用：10 ~ 60岁正常人血清中甲氧基丙烯酸含量为71.8 12.3 mg/l，无年龄、性别差异，肝功能不全及慢性肾功能衰竭患者各疾病血清中甲氧基丙烯酸含量较低。值得注意的是，在急性肝炎恢复期和药物性肝炎恢复期，该值较高。这些结果表明，该蛋白主要在肝脏中产生，测定该蛋白对评价肝实质细胞损伤及其修复有重要意义。本项目也促进了对低分子蛋白质的研究。少

项目成果

Y.Itoh.: Protides in biologocal fluids. 35. 269-272 (1987)

Y.Itoh.：生物体液中的蛋白质。

伊藤喜久: 臨床化学年会記録. 27. 66-69 (1987)

伊藤义久：临床化学年会记录 27. 66-69 (1987)

Y.Itoh.: Contri.Nepirolo.68. 172-178 (1988)

Y.Itoh.：Contri.Nepirolo.68。

Y. Itoh: "A latex agglutination test for urinary _2-microglobulin." Contri. Nephrolo.68. 172-178 (1988)

Y. Itoh：“尿液 _2-微球蛋白的乳胶凝集试验。”

伊藤喜久: 臨床化学. (1988)

伊藤义久：临床化学（1988）。