Redesign of a Copper Storage Protein into an Artificial Hydrogenase

Dhanashree Selvan, Pallavi Prasad, Erik R. Farquhar, Yelu Shi, Skyler Crane, Yong Zhang, Saumen Chakraborty

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

We report the construction of an artificial hydrogenase (ArH) by reengineering a Cu storage protein (Csp1) into a Ni-binding protein (NBP) employing rational metalloprotein design. The hypothesis driven design approach involved deleting existing Cu sites of Csp1 and identification of a target tetrathiolate Ni binding site within the protein scaffold followed by repacking the hydrophobic core. Guided by modeling, the NBP was expressed and purified in high purity. NBP is a well-folded and stable construct displaying native-like unfolding behavior. Spectroscopic and computational studies indicated that NBP bound nickel in a distorted square planar geometry that validated the design. Ni(II)-NBP is active for photoinduced H2 evolution following a reductive quenching mechanism. Ni(II)-NBP catalyzed H+ reduction to H2 gas electrochemically as well. Analysis of the catalytic voltammograms established a proton-coupled electron transfer mechanism. Electrolysis studies confirmed H2 evolution with quantitative Faradaic yields. Our studies demonstrate an important scope of rational metalloprotein design that allows imparting functions into protein scaffolds that have natively not evolved to possess the same function of the target metalloprotein constructs.

Original languageEnglish
Pages (from-to)5847-5859
Number of pages13
JournalACS Catalysis
Volume9
Issue number7
DOIs
StatePublished - 5 Jul 2019

Keywords

  • artificial metalloenzymes
  • catalysis
  • hydrogenases
  • protein design
  • spectroscopy

Fingerprint

Dive into the research topics of 'Redesign of a Copper Storage Protein into an Artificial Hydrogenase'. Together they form a unique fingerprint.

Cite this