Issue 39, 2017

Spectroscopic evidence for cofactor–substrate interaction in the radical-SAM enzyme TYW1

Abstract

TYW1 is a metalloenzyme involved in the modifications of guanosine 37 of Phe-tRNA of Eukaryota and Archaea. It catalyzes the second step of Wybutosine biosynthesis, which consists of the formation of the tricyclic compound imG-14 from m1G using pyruvate and SAM (S-adenosyl-methionine) as co-substrates. Two [4Fe–4S] clusters are needed in the catalytic process. One effects the reductive binding of SAM, which initiates the radical reaction that inserts a C–C moiety into m1G. The other [4Fe–4S] cluster binds the pyruvate molecule that provides the C–C motif. Using a combination of EPR and Mössbauer spectroscopy, we have been able to probe the binding of both cofactors to the FeS clusters. The results highlight an interaction between pyruvate and SAM, indicating that they bind in close vicinity inside the catalytic pocket. They also indicate a chelating binding mode of pyruvate to the accessible Fe site of the corresponding FeS cluster. This binding mode has been used to construct a docking model of holoTYW1 with pyruvate and SAM, which is consistent with the spectroscopic findings.

Graphical abstract: Spectroscopic evidence for cofactor–substrate interaction in the radical-SAM enzyme TYW1

Supplementary files

Article information

Article type
Paper
Submitted
28 фев 2017
Accepted
05 июн 2017
First published
05 июн 2017

Dalton Trans., 2017,46, 13211-13219

Spectroscopic evidence for cofactor–substrate interaction in the radical-SAM enzyme TYW1

V. Kathirvelu, P. Perche-Letuvée, J. Latour, M. Atta, F. Forouhar, S. Gambarelli and R. Garcia-Serres, Dalton Trans., 2017, 46, 13211 DOI: 10.1039/C7DT00736A

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements