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![Protein Film Infrared Electrochemistry Demonstrated for Study of H2 Oxidation by a [NiFe] Hydrogenase](/_next/image?url=https%3A%2F%2Fcloudfront.jove.com%2FCDNSource%2Fteasers%2F55858.jpg&w=3840&q=50)
Protein Film Infrared Electrochemistry Demonstrated for Study of H2 Oxidation by a [NiFe] Hydrogenase
Published on: December 4, 2017
Biosynthesis of the Selenium-Substituted [FeFe]-Hydrogenases
Xin Yu1, Toby Woods1, Yu Zhang1
1School of Chemical Sciences, University of Illinois, Urbana, Illinois 61801, United States.
None:
The [FeFe]-hydrogenase from Chlamydomonas reinhardtii (CrHydA1) has been prepared with Se in place of S at the [2Fe]H active site by HydF-mediated maturation using [HFe2(μ-SeH)(μ-Se)(CN)2(CO)4]2- ([2]2-) and the apoenzyme. Complex [2]2- is a hydride─a rare case where a metalloselenol and the (naturally occurring) metallothiol [Fe2(μ-SH)2(CN)2(CO)4]2-─adopt different structures. The structure of [2]2- was deduced based on 1H and 77Se NMR spectroscopy. From [2]2-, highly active CrHydA1-Se2 can be efficiently produced using CH2O, but not serine, as the precursor to the azadiselenolate cofactor. EPR/ENDOR spectroscopic studies were corroborated with isotopically pure CrHydA1-77Se2 produced by maturation with [Fe2[(μ-77SeCH2)2NH](CN)2(CO)4]2-. CrHydA1-Se2 and native CrHydA1 have distinct, but quite similar geometric and electronic structures and redox properties.
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