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通过肿瘤细胞SELEX鉴定的肌腱蛋白-C适配体:指数富集配体的系统进化

A tenascin-C aptamer identified by tumor cell SELEX: systematic evolution of ligands by exponential enrichment.

作者信息

Daniels Dion A, Chen Hang, Hicke Brian J, Swiderek Kristine M, Gold Larry

机构信息

Gene Expression and Protein Biochemistry, GlaxoSmithKline Medicines Research Centre, Stevenage, Hertfordshire SG1 2NY, England.

出版信息

Proc Natl Acad Sci U S A. 2003 Dec 23;100(26):15416-21. doi: 10.1073/pnas.2136683100. Epub 2003 Dec 15.

Abstract

The targeting of molecular repertoires to complex systems rather than biochemically pure entities is an accessible approach that can identify proteins of biological interest. We have probed antigens presented by a monolayer of tumor cells for their ability to interact with a pool of aptamers. A glioblastoma-derived cell line, U251, was used as the target for systematic evolution of ligands by exponential enrichment by using a single-stranded DNA library. We isolated specifically interacting oligonucleotides, and biochemical strategies were used to identify the protein target for one of the aptamers. Here we characterize the interaction of the DNA aptamer, GBI-10, with tenascin-C, an extracellular protein found in the tumor matrix. Tenascin-C is believed to be involved in both embryogenesis and oncogenesis pathways. Systematic evolution of ligands by exponential enrichment appears to be a successful strategy for the a priori identification of targets of biological interest within complex systems.

摘要

将分子文库靶向复杂系统而非生物化学纯实体是一种可行的方法,它能够识别具有生物学意义的蛋白质。我们已探究单层肿瘤细胞所呈递的抗原与一组适配体相互作用的能力。通过使用单链DNA文库,源自胶质母细胞瘤的细胞系U251被用作通过指数富集进行配体系统进化的靶标。我们分离出了特异性相互作用的寡核苷酸,并运用生化策略鉴定了其中一种适配体的蛋白质靶标。在此,我们描述了DNA适配体GBI-10与腱生蛋白-C(一种在肿瘤基质中发现的细胞外蛋白)之间的相互作用。腱生蛋白-C被认为参与胚胎发生和肿瘤发生途径。通过指数富集进行配体系统进化似乎是在复杂系统中先验鉴定具有生物学意义的靶标的成功策略。

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