JCVI: A Broad Survey Reveals Substitution Tolerance of Residues Ligating FeS Clusters in [NiFe] Hydrogenase.
 
 
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Citation

Yonemoto IT, Clarkson BN, Smith HO, Weyman PD

A Broad Survey Reveals Substitution Tolerance of Residues Ligating FeS Clusters in [NiFe] Hydrogenase.

BMC Biochemistry. 2014 Jun 17; 15. : 10.

External Citation

Abstract

In order to understand the effects of FeS cluster attachment in [NiFe] hydrogenase, we undertook a study to substitute all 12 amino acid positions normally ligating the three FeS clusters in the hydrogenase small subunit. Using the hydrogenase from Alteromonas macleodii "deep ecotype" as a model, we substituted one of four amino acids (Asp, His, Asn, Gln) at each of the 12 ligating positions because these amino acids are alternative coordinating residues in otherwise conserved-cysteine positions found in a broad survey of NiFe hydrogenase sequences. We also hoped to discover an enzyme with elevated hydrogen evolution activity relative to a previously reported "G1" (H230C/P285C) improved enzyme in which the medial FeS cluster Pro and the distal FeS cluster His were each substituted for Cys.