Intracellular transport of Mutant alpha_2-plasmin inhibitor

突变型 α_2-纤溶酶抑制剂的细胞内转运

• 批准号: 03454522
• 负责人: AOKI Nobuo
• 金额: $ 4.22万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-03454522/
• 关键词: alpha_2-plasmin inhibitor; Congenital Deficiency; Intracellular Transport; Gene Mutation; Impairement of Secretion; 細胞内輸送障害; 失天性欠損症; アルフア2プラスミンインヒビタ-; 欠損症; α_2PIーOkinawa

alpha_2-Plasmin inhibitor (alpha_2PI) deficiency Okinawa results from defective secretion of the inhibitor from the liver and appears to be a direct consequence of the deletion of Glu^<137> in the amino acid sequence of alpha_2PI.To examine the effects of replacing the amino acid occupying position 137 or deleting its neighboring amino acid on alpha_2PI secretion, we used oligonucleotide-directed mutagenesis of alpha_2PI cDNA to change the codon specifying Glu^<137> or delete a codon specifying its neighboring amino acid. The effects were determined by pulsechase experiments and by enzyme-linked immunosorbent assay of media from transiently transfected COS-7 cells. Replacement of Glu^<137> with an amino acid other than Cys had little effect on alpha_2PI secretion. In contrast, deletion of an amino acid in a region spanning a sequence of less than 30 amino acids including positions 127 and 137 severely impaired the secretion. The results suggest that structural integrity of the region, rather than its component amino acids, is important for the intracellular transport and secretion of alpha_2PI.

Alpha_2-纤溶酶抑制物(α_2pi)缺乏症是由肝脏分泌抑制物缺陷引起的，似乎是Alpha_2pi氨基酸序列中Glu^&lt；137&gt；缺失的直接结果。为了研究取代137位氨基酸或删除其邻近氨基酸对Alpha_2pi分泌的影响，我们利用α_2pi基因的寡核苷酸定向突变来改变指定Glu^&lt；137&gt；的密码子或删除指定其邻近氨基酸的密码子。通过脉冲期实验和瞬时转基因COS-7细胞培养上清液的酶联免疫吸附试验来确定其效果。用Cys以外的氨基酸取代Glu^&lt；137&gt；对α_2pi的分泌几乎没有影响。相反，在127和137位的少于30个氨基酸的序列中缺失一个氨基酸会严重损害分泌。结果表明，该区域的结构完整性，而不是其组成的氨基酸，对细胞内α2pi的运输和分泌是重要的。

项目成果

Takatoshi Koyama: "Relationship between postーtranslational glycosylation and anticoagulant function of secretable recanbinant mutants of human thrambomodulin" British Journal of Haematology. 78. 515-522 (1991)

Takatoshi Koyama：“人血栓调节蛋白分泌型重组突变体的翻译后糖基化与抗凝功能之间的关系”英国血液学杂志 78. 515-522 (1991)。

Nobuo Aoki: "alpha 2 plasmin inhibitor and its clinical significance" Asian Medical Journal. 36. 557-565 (1993)

青木伸夫：《α2纤溶酶抑制剂及其临床意义》亚洲医学杂志。

Atsushi Kato et al.: "Localization of the human α_2-plasmin inhibitor gene(PLI) to 17p13" Cytogenetics and Cell genetics. 62. 190-191 (1993)

Atsushi Kato 等人：“人类 α_2-纤溶酶抑制剂基因 (PLI) 定位于 17p13”《细胞遗传学和细胞遗传学》62. 190-191 (1993)。

Shigeo Toyota et al.: "Secretion of α_2-plasmin inhibitor is imfaired by amino acid deletion in a small region of the molecule" Journal of Biochemistry. 115. 293-297 (1994)

Shigeo Toyota 等人：“α_2-纤溶酶抑制剂的分泌因分子小区域的氨基酸缺失而受到损害”《生物化学杂志》115. 293-297 (1994)。

Koyama,Takatoshi: "Purification of human α_2-plasmin inhibitor by a one-step immunoaffinity chromatography procedure" Thrombosis Research. 66. 451-454 (1992)

Koyama，Takatoshi：“通过一步免疫亲和层析程序纯化人 α_2-纤溶酶抑制剂”《血栓形成研究》66. 451-454 (1992)。