H/D exchange reactions and mechanistic aspects of the hydrogenases

✍ Scribed by Paulette M. Vignais

• Publisher: Elsevier Science
• Year: 2005
• Tongue: English
• Weight: 296 KB
• Volume: 249
• Category: Article
• ISSN: 0010-8545
• DOI: 10.1016/j.ccr.2005.01.026

✦ Synopsis

In the presence of D 2 gas (in a D 2 /H 2 O system), hydrogenases catalyze the proton-deuterium (H/D) exchange reaction, which results in the formation of HD and H 2 , measurable by mass spectrometry at mass peaks m/e 3 and 2, respectively. This technique can be used to study the activity and function of hydrogenases both in vitro, with isolated enzymes, and in vivo, with whole cells. It allows to follow the reductive activation of oxidized [NiFe]-hydrogenases and to identify physiological activators such as NADH and NADPH, of bidirectional hydrogenases. Besides, since the H/D exchange reaction involves proton exchange with the solvent, it has been used to monitor the effects of site-directed mutagenesis of amino acid residues postulated to belong to proton channels.