Issue 44, 2019

Revealing the transient conformations of a single flavin adenine dinucleotide using an aerolysin nanopore

Abstract

Flavin adenine dinucleotide (FAD) as a cofactor is involved in numerous important metabolic pathways where the biological function is intrinsically related to its transient conformations. The confined space of enzymes requires FAD set in its specific intermediate conformation. However, conventional methods only detect stable conformations of FAD molecules, while transient intermediates are hidden in ensemble measurements. There still exists a challenge to uncover the transient conformation of each FAD molecule, which hinders the understanding of the structure–activity relationship of the FAD mechanism. Here, we employ the electrochemically confined space of an aerolysin nanopore to directly characterize a series of transient conformations of every individual FAD. Based on distinguishable current blockages, the “stack”, “open”, and four quasi-stacked FADs are clearly determined in solution, which is further confirmed by temperature-dependent experiments and mutant aerolysin assay. Combined with molecular dynamics simulations, we achieved a direct correlation between the residual current ratio (I/I0) and FAD backbone angle. These results would facilitate further understanding of the structure–activity relationship in the flavoprotein.

Graphical abstract: Revealing the transient conformations of a single flavin adenine dinucleotide using an aerolysin nanopore

Supplementary files

Article information

Article type
Edge Article
Submitted
27 jun. 2019
Accepted
20 sep. 2019
First published
23 sep. 2019
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY-NC license

Chem. Sci., 2019,10, 10400-10404

Revealing the transient conformations of a single flavin adenine dinucleotide using an aerolysin nanopore

M. Li, Y. Wang, Y. Ying and Y. Long, Chem. Sci., 2019, 10, 10400 DOI: 10.1039/C9SC03163D

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