Escherichia coli biotin synthase produces selenobiotin. Further evidence of the involvement of the [2Fe-2S]2+ cluster in the sulfur insertion step

Biochemistry
Bernadette Tse Sum BuiA Marquet

Abstract

Biotin synthase, a member of the "radical SAM" family, catalyzes the final step of the biotin biosynthetic pathway, namely, the insertion of a sulfur atom into dethiobiotin. The as-isolated enzyme contains a [2Fe-2S](2+) cluster, but the active enzyme requires an additional [4Fe-4S](2+) cluster, which is formed in the presence of Fe(NH(4))(2)(SO(4))(2) and Na(2)S in the in vitro assay. The role of the [4Fe-4S](2+) cluster is to mediate the electron transfer to SAM, while the [2Fe-2S](2+) cluster is involved in the sulfur insertion step. To investigate the selenium version of the reaction, we have depleted the enzyme of its iron and sulfur and reconstituted the resulting apoprotein with FeCl(3) and Na(2)Se to yield a [2Fe-2Se](2+) cluster. This enzyme was assayed in vitro with Na(2)Se in place of Na(2)S to enable the formation of a [4Fe-4Se](2+) cluster. Selenobiotin was produced, but the activity was lower than that of the as-isolated [2Fe-2S](2+) enzyme in the presence of Na(2)S. The [2Fe-2Se](2+) enzyme was additionally assayed with Na(2)S, to reconstitute a [4Fe-4S](2+) cluster, in case the latter was more efficient than a [4Fe-4Se](2+) cluster for the electron transfer. Indeed, the activity was improved, but in that case, a...Continue Reading

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Citations

Sep 28, 2007·Natural Product Reports·Andrée MarquetMartin J Warren
Mar 26, 2011·Molecular BioSystems·Steven Lin, John E Cronan
Apr 2, 2013·Nature Chemical Biology·Farhad ForouharMarc Fontecave
Feb 13, 2016·Journal of the American Chemical Society·Bradley J Landgraf, Squire J Booker
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Aug 1, 2012·Biochimica Et Biophysica Acta·Nicholas D Lanz, Squire J Booker
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Jan 31, 2014·Chemical Reviews·Joan B BroderickEric M Shepard
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Sep 16, 2015·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Atanu RanaAbhishek Dey
Dec 25, 2007·Archives of Biochemistry and Biophysics·Michael R ReydaJoseph T Jarrett
Nov 28, 2017·Dalton Transactions : an International Journal of Inorganic Chemistry·L KertessU-P Apfel
Jan 12, 2021·FEMS Microbiology Reviews·Chaiyos Sirithanakorn, John E Cronan

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