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人T细胞系Jurkat中白细胞介素2信使核糖核酸和蛋白质的动力学及环孢菌素A的作用

Kinetics of interleukin 2 mRNA and protein produced in the human T-cell line Jurkat and effect of cyclosporin A.

作者信息

Nordmann R, Andersen E, Trussardi R, Mazer N A

机构信息

Department of Preclinical Research, Sandoz Ltd., Basle, Switzerland.

出版信息

Biochemistry. 1989 Feb 21;28(4):1791-7. doi: 10.1021/bi00430a055.

Abstract

The kinetics of interleukin 2 mRNA accumulation in the leukemic T-cell line Jurkat, which can be induced with phytohemagglutinin and phorbol 12-myristate 13-acetate to produce large amounts of interleukin 2, was analyzed by a modified DNA-excess solution hybridization assay using a 5'-32P-labeled oligodeoxyribonucleotide 30 bases long as probe. Cyclosporin A was used as a valuable tool to gain more insight into the quantitative aspects of interleukin 2 production, on the basis of the assumption that transcription of the interleukin 2 gene is completely inhibited shortly after administration of cyclosporin A. The half-life of interleukin 2 mRNA was estimated to be approximately 2 h. With the aid of simple mathematical models, we have been able to relate the concentration of interleukin 2 protein in the supernatant to the interleukin 2 mRNA kinetics. This novel quantitative kinetic analysis revealed that, independent of the absence or presence of cyclosporin A, interleukin 2 protein is synthesized at a rate of approximately 1.3 molecules per molecule of interleukin 2 mRNA per second and secreted within 2 h after it is synthesized and that its half-life in the supernatant is approximately 10 h.

摘要

白血病T细胞系Jurkat可被植物血凝素和佛波酯诱导产生大量白细胞介素2,利用一种改良的DNA过量溶液杂交试验,以一条30个碱基长的5'-32P标记的寡脱氧核糖核苷酸作为探针,分析了该细胞系中白细胞介素2 mRNA积累的动力学。基于环孢素A给药后不久白细胞介素2基因转录被完全抑制这一假设,将环孢素A用作一种有价值的工具,以更深入了解白细胞介素2产生的定量方面。白细胞介素2 mRNA的半衰期估计约为2小时。借助简单的数学模型,我们能够将上清液中白细胞介素2蛋白的浓度与白细胞介素2 mRNA动力学联系起来。这种新颖的定量动力学分析表明,无论有无环孢素A,白细胞介素2蛋白的合成速率约为每秒每分子白细胞介素2 mRNA 1.3个分子,并在合成后2小时内分泌,其在上清液中的半衰期约为10小时。

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