Wano Y, Uchiyama T, Yodoi J, Uchino H
Microbiol Immunol. 1985;29(5):451-66. doi: 10.1111/j.1348-0421.1985.tb00846.x.
Biosynthetic processing of the T-cell surface receptor for interleukin-2 was investigated in a cultured human T-cell line MT-1 by means of metabolic and cell surface radiolabeling followed by immunoprecipitation with a monoclonal anti-receptor antibody (anti-Tac) and analysis by one- and two-dimensional polyacrylamide gel electrophoresis. The nascent precursor of the receptor (Mr = about 40,000, pI = 6.2-6.5) underwent a post-translational modification giving rise to the mature receptor (IL-2R; Mr = 60,000-65,000, pI = 4.2-4.7) within 2-4 hr. The post-translational processing of IL-2R caused a 20,000-25,000 increase in apparent molecular weight and a 2.0-2.5 acidic shift in the isoelectric point. The increase in molecular weight was attributable mainly to addition of sugar residues including glucosamine and galactose, and the charge shift to the addition of sialic acids. A carboxylic ionophore monensin completely blocked the maturation of IL-2R at the mid-stage of the processing. Fatty acid attachment appeared to comprise one of the steps of the post-translational modification. Two-dimensional analyses of IL-2R biosynthesis enabled identification of the precursor of IL-2R and its intermediate forms, from which it was partially possible to estimate reactions involved in the maturation of the precursor molecule.
利用代谢和细胞表面放射性标记，随后用单克隆抗受体抗体（抗 Tac）进行免疫沉淀，并通过一维和二维聚丙烯酰胺凝胶电泳分析，在培养的人 T 细胞系 MT-1 中研究了白细胞介素-2 的 T 细胞表面受体的生物合成过程。受体的新生前体（Mr = 约 40,000，pI = 6.2 - 6.5）经历了翻译后修饰，在 2 - 4 小时内产生成熟受体（IL-2R；Mr = 60,000 - 65,000，pI = 4.2 - 4.7）。IL-2R 的翻译后加工导致表观分子量增加 20,000 - 25,000，等电点向酸性方向移动 2.0 - 2.5。分子量的增加主要归因于包括氨基葡萄糖和半乳糖在内的糖残基的添加，电荷的移动归因于唾液酸的添加。羧酸离子载体莫能菌素在加工的中期完全阻断了 IL-2R 的成熟。脂肪酸附着似乎是翻译后修饰的步骤之一。对 IL-2R 生物合成的二维分析能够鉴定 IL-2R 的前体及其中间形式，从中部分地可以估计前体分子成熟过程中涉及的反应。
