Purification of the proline-rich homeodomain protein
Purification of the proline-rich homeodomain protein
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DOI：
10.1016/s1570-0232(02)00740-7
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发表时间：
2003-03-25
影响因子：
3
通讯作者：
Jayaraman, PS
中科院分区：
文献类型：
--
作者：
Butcher, AJ;Gaston, K;Jayaraman, PS
富含脯氨酸的同源结构域蛋白 (PRH)，也称为 Hex，是一种在多种细胞类型中表达的转录抑制蛋白。 PRH 蛋白包含一个富含脯氨酸的 N 端结构域（当连接到异源 DNA 结合结构域时可以抑制转录）、一个介导序列特异性 DNA 结合的中心同源结构域以及一个功能未知的酸性 C 端结构域。尽管 PRH 的各个结构域已在细菌细胞中表达为 GST 和组氨酸标记的融合蛋白，但表达和纯化全长蛋白的尝试却收效甚微。在这里，我们描述了组氨酸标记的全长 PRH 融合蛋白的纯化。这里描述的蛋白质将使我们能够确定 PRH 抑制转录的机制。 (C) 2002 Elsevier Science B.V. 保留所有权利。
The proline-rich homeodomain protein (PRH), also known as Hex, is a transcriptional repressor expressed in a variety of cell types. The PRH protein contains a proline-rich N-terminal domain that can repress transcription when attached to a heterologous DNA binding domain, a central homeodomain that mediates sequence-specific DNA binding, and an acidic C-terminal domain of unknown function. Although individual domains of PRH have been expressed in bacterial cells as GST- and histidine-tagged fusion proteins, attempts to express and purify the full-length protein have met with little success. Here we describe the purification of a histidine-tagged full-length PRH fusion protein. The protein described here will allow us to determine the mechanisms whereby PRH represses transcription. (C) 2002 Elsevier Science B.V. All rights reserved.