Photocatalytic Hydrogen Evolution by a De Novo Designed Metalloprotein that Undergoes Ni-Mediated Oligomerization Shift

De novo metalloprotein design involves the construction of proteins guided by specific repeat patterns of polar and apolar residues, which, upon self-assembly, provide a suitable environment to bind metals and produce artificial metalloenzymes. While a wide range of functionalities have been realized in de novo designed metalloproteins, the functional repertoire of such constructs towards alternative energy-relevant catalysis is currently limited. Here we show the application of de novo approach to design a functional H-2 evolving protein. The design involved the assembly of an amphiphilic peptide featuring cysteines at tandem a/d sites of each helix. Intriguingly, upon Ni-II addition, the oligomers shift from a major trimeric assembly to a mix of dimers and trimers. The metalloprotein produced H-2 photocatalytically with a bell-shape pH dependence, having a maximum activity at pH 5.5. Transient absorption spectroscopy is used to determine the timescales of electron transfer as a function of pH. Selective outer sphere mutations are made to probe how the local environment tunes activity. A preferential enhancement of activity is observed via steric modulation above the Ni-II site, towards the N-termini, compared to below the Ni-II site towards the C-termini.

Automated summary

This article introduces a new method for protein design, which involves constructing proteins with specific repeat patterns of polar and apolar residues to create a suitable environment for binding metals and producing artificial metalloenzymes. The designed protein successfully achieved hydrogen evolution and has potential applications in alternative energy-relevant catalysis.