4.1 Article

Importance of the MbtH-like protein TioT for production and activation of the thiocoraline adenylation domain of TioK

期刊

MEDCHEMCOMM
卷 3, 期 8, 页码 950-955

出版社

ROYAL SOC CHEMISTRY
DOI: 10.1039/c2md20131c

关键词

-

资金

  1. Life Sciences Institute
  2. College of Pharmacy at the University of Michigan
  3. NSF [MCB 1149427]
  4. Vahlteich Research Award

向作者/读者索取更多资源

The 3-hydroxyquinaldic acid (3HQA) chromophores of thiocoraline are essential for the biological DNA bisintercalating function of this antitumor agent. The 3HQA units are also proposed to play a critical role in the resistance mechanism of the thiocoraline-producing organism against this natural product. Because of their important functions, there is a great interest in understanding the 3HQA formation from L-Trp. The first proposed committed steps during 3HQA biosynthesis consist of conversion of L-Trp into L-Trp-AMP by the adenylation domain of TioK followed by installation of the activated amino acid onto the thiolation domain of this didomain enzyme. However, testing this series of events has been hindered by the inability to heterologously express soluble TioK. Here, we demonstrated that the MbtH-like protein TioT is required for production and activation of TioK. With soluble functional TioK in hand, we established the amino acid substrate profile and kinetically characterized this enzyme. By site-directed mutagenesis of TioT, we also investigated the significance of three Pro residues that are universally conserved in MbtH-like proteins.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.1
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据