4.7 Article

Spatial Structure of NanoFAST in the Apo State and in Complex with its Fluorogen HBR-DOM2

期刊

出版社

MDPI
DOI: 10.3390/ijms231911361

关键词

fluorogen-activating protein; FAST; nanoFAST; spatial structure; dynamics; ligand specificity; fluorogen; binding constant; structure

资金

  1. Russian Science Foundation [18-73-10105]

向作者/读者索取更多资源

NanoFAST is a fluorogen-activating protein and the deletion of the N-terminus of FAST destabilizes the C-terminal domain in the apo state, affecting ligand binding propensity. The structure of the nanoFAST/HBR-DOM2 complex reveals atomistic details of nanoFAST interactions with ligands and explains ligand specificity.
NanoFAST is a fluorogen-activating protein and can be considered one of the smallest encodable fluorescent tags. Being a shortened variant of another fluorescent tag, FAST, nanoFAST works nicely only with one out of all known FAST ligands. This substantially limits the applicability of this protein. To find the reason for such a behavior, we investigated the spatial structure and dynamics of nanoFAST, both in the apo state and in the complex with its fluorogen molecule, using the solution NMR spectroscopy. We showed that the truncation of FAST did not affect the structure of the remaining part of the protein. Our data suggest that the deleted N-terminus of FAST destabilizes the C-terminal domain in the apo state. While it does not contact the fluorogen directly, it serves as a free energy reservoir that enhances the ligand binding propensity of the protein. The structure of nanoFAST/HBR-DOM2 complex reveals the atomistic details of nanoFAST interactions with the rhodanine-based ligands and explains the ligand specificity. NanoFAST selects ligands with the lowest dissociation constants, 2,5-disubstituted 4-hydroxybenzyldienerhodainines, which allow the non-canonical intermolecular CH-N hydrogen bonding and provide the optimal packing of the ligand within the hydrophobic cavity of the protein.

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

4.7
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