Development and application of fully functional epitope-tagged forms of transforming growth factor-β
Development and application of fully functional epitope-tagged forms of transforming growth factor-β
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DOI：
10.1016/s0022-1759(02)00090-x
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发表时间：
2002-08-01
影响因子：
2.2
通讯作者：
Letterio, JJ
中科院分区：
文献类型：
--
作者：
Wolfraim, LA;Alkemade, GM;Letterio, JJ
转化生长因子- β (tgf - β)已被发现在各种小鼠疾病模型中具有治疗益处，并具有潜在的临床应用价值。然而，追踪外源性给药的分布的能力，这些蛋白质的重组形式受到内源性tgf - β和相关tgf - β亚型的交叉反应性的限制。我们描述了新的FLAG和血凝素（HA）标记的成熟tgf - β - 1，它们保留了充分的生物活性，这被证明它们能够抑制Mv1Lu上皮细胞的生长，并诱导tgf - β信号传导介质smad2的磷酸化。共聚焦免疫荧光显微镜可检测转染细胞内的FLAG-和ha - tgf - β 1。我们还描述了设计用于特异性检测表位标记的tgf - β的夹心elisa，并通过流式细胞术证明了这些标记配体作为tgf - β受体表达探针的效用。这些全功能表位标记的tgf - β蛋白的设计将促进体内肽药效学评估和tgf - β配体受体复合物运输等研究。(C) 2002年Elsevier Science B.V.出版
Administration of transforming growth factor-beta (TGF-beta) has been found to be of therapeutic benefit in various mouse disease models and has potential clinical usefulness. However, the ability to track the distribution of exogenously administered, recombinant forms of these proteins has been restricted by cross-reactivity with endogenous TGF-beta and related TGF-beta isoforms. We describe novel FLAG- and hemagglutinin (HA)-tagged versions of mature TGF-beta1 that retain full biological activity as demonstrated by their ability to inhibit the growth of Mv1Lu epithelial cells, and to induce phosphorylation of the TGF-beta signaling intermediate, smad 2. Intracellular FLAG- and HA-TGF-beta1 can be detected in transfected cells by confocal immunofluorescence microscopy. We also describe sandwich ELISAs designed to specifically detect epitope-tagged TGF-beta and demonstrate the utility of these tagged ligands as probes for TGF-beta receptor expression by flow cytometry. The design of these fully functional epitope-tagged TGF-beta proteins should facilitate studies such as the evaluation of in vivo peptide pharmacodynamics and trafficking of TGF-beta ligand-receptor complexes. (C) 2002 Published by Elsevier Science B.V.