...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Scientific reports. >Passenger sequences can promote interlaced dimers in a common variant of the maltose-binding protein
【24h】

Passenger sequences can promote interlaced dimers in a common variant of the maltose-binding protein

机译：乘客序列可以在麦芽糖结合蛋白的常见变体中促进相互扫描的二聚体

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The maltose-binding protein (MBP) is one of the most frequently used protein tags due to its capacity to stabilize, solubilize and even crystallize recombinant proteins that are fused to it. Given that MBP is thought to be a highly stable monomeric protein with known characteristics, fused passenger proteins are often studied without being cleaved from MBP. Here we report that a commonly used engineered MBP version (mutated to lower its surface entropy) can form interlaced dimers when fused to short protein sequences derived from the focal adhesion kinase (FAK) or the homologous protein tyrosine kinase 2 (PYK2). These MBP dimers still bind maltose and can interconvert with monomeric forms in vitro under standard conditions despite a contact surface of more than 11,000 ?sup2/sup. We demonstrate that both the mutations in MBP and the fused protein sequences were required for dimer formation. The FAK and PYK2 sequences are less than 40% identical, monomeric, and did not show specific interactions with MBP, suggesting that a variety of sequences can promote this MBP dimerization. MBP dimerization was abrogated by reverting two of the eight mutations introduced in the engineered MBP. Our results provide an extreme example for induced reversible domain-swapping, with implications for protein folding dynamics. Our observations caution that passenger-promoted MBP dimerization might mislead experimental characterization of the fused protein sequences, but also suggest a simple mutation to stop this phenomenon.
机译：麦芽糖结合蛋白（MBP）是由于其稳定，溶解和甚至结晶融合到其的重组蛋白质，最常使用的蛋白质标签之一。鉴于MBP被认为是具有已知特征的高度稳定的单体蛋白质，通常研究融合的乘客蛋白而不从MBP切割。在这里，我们报告说，当融合到源自焦粘连激酶（Fak）或同源蛋白酪氨酸激酶2（Pyk2）的短蛋白序列融合时，常用的工程MBP版本（突变为降低其表面熵）可以形成隔行二聚体。这些MBP二聚体仍然结合麦芽糖，并且尽管接触表面超过11,000μl，但可以在标准条件下在体外与单体形式相互转换。我们证明，二聚体形成需要MBP中的突变和融合蛋白序列。 FAK和PyK2序列均不到40％相同的单体，并且没有显示与MBP的特异性相互作用，表明各种序列可以促进该MBP二聚化。通过在工程化MBP中介绍八个突变中的两个突变来消除MBP二聚化。我们的结果为诱导可逆域交换提供了一个极端的例子，具有对蛋白质折叠动态的影响。我们的观察员小心乘用MBP二聚化可能误导融合蛋白质序列的实验表征，但也表明了一种抑制这种现象的简单突变。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号