Issue 18, 2019

Monitoring H-cluster assembly using a semi-synthetic HydF protein


The [FeFe] hydrogenase enzyme interconverts protons and molecular hydrogen with remarkable efficiency. The reaction is catalysed by a unique metallo-cofactor denoted as the H-cluster containing an organometallic dinuclear Fe component, the [2Fe] subsite. The HydF protein delivers a precursor of the [2Fe] subsite to the apo-[FeFe] hydrogenase, thus completing the H-cluster and activating the enzyme. Herein we generate a semi-synthetic form of HydF by loading it with a synthetic low valent dinuclear Fe complex. We show that this semi-synthetic protein is practically indistinguishable from the native protein, and utilize this form of HydF to explore the mechanism of H-cluster assembly. More specifically, we show that transfer of the precatalyst from HydF to the hydrogenase enzyme results in the release of CO, underscoring that the pre-catalyst is a four CO species when bound to HydF. Moreover, we propose that an electron transfer reaction occurs during H-cluster assembly, resulting in an oxidation of the [2Fe] subsite with concomitant reduction of the [4Fe4S] cluster present on the HydF protein.

Graphical abstract: Monitoring H-cluster assembly using a semi-synthetic HydF protein

Supplementary files

Article information

Article type
26 Oct 2018
21 Dec 2018
First published
03 Jan 2019
This article is Open Access
Creative Commons BY license

Dalton Trans., 2019,48, 5978-5986

Monitoring H-cluster assembly using a semi-synthetic HydF protein

B. Németh, C. Esmieu, H. J. Redman and G. Berggren, Dalton Trans., 2019, 48, 5978 DOI: 10.1039/C8DT04294B

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