Catalysis-dependent selenium incorporation and migration in the nitrogenase active site iron-molybdenum cofactor

Spatzal, Thomas; Perez, Kathryn A.; Howard, James B.; Rees, Douglas C.

doi:10.7554/elife.11620

Title: Catalysis-dependent selenium incorporation and migration in the nitrogenase active site iron-molybdenum cofactor

Journal Article · Wed Dec 16 00:00:00 EST 2015 · eLife

DOI:https://doi.org/10.7554/elife.11620· OSTI ID:1628840

Spatzal, Thomas ^[1]; Perez, Kathryn A. ^[2]; Howard, James B. ^[3]; Rees, Douglas C. ^[1]

California Institute of Technology (CalTech), Pasadena, CA (United States). Howard Hughes Medical Inst.; California Institute of Technology (CalTech), Pasadena, CA (United States). Division of Chemistry and Chemical Engineering
California Institute of Technology (CalTech), Pasadena, CA (United States). Division of Chemistry and Chemical Engineering
California Institute of Technology (CalTech), Pasadena, CA (United States). Division of Chemistry and Chemical Engineering; Univ. of Minnesota, Minneapolis, MN (United States). Dept. of Biochemistry, Molecular Biology and Biophysics

Dinitrogen reduction in the biological nitrogen cycle is catalyzed by nitrogenase, a two-component metalloenzyme. Understanding of the transformation of the inert resting state of the active site FeMo-cofactor into an activated state capable of reducing dinitrogen remains elusive. Here we report the catalysis dependent, site-selective incorporation of selenium into the FeMo-cofactor from selenocyanate as a newly identified substrate and inhibitor. The 1.60 Å resolution structure reveals selenium occupying the S2B site of FeMo-cofactor in the Azotobacter vinelandii MoFe-protein, a position that was recently identified as the CO-binding site. The Se2B-labeled enzyme retains substrate reduction activity and marks the starting point for a crystallographic pulse-chase experiment of the active site during turnover. Through a series of crystal structures obtained at resolutions of 1.32–1.66 Å, including the CO-inhibited form of Av1-Se2B, the exchangeability of all three belt-sulfur sites is demonstrated, providing direct insights into unforeseen rearrangements of the metal center during catalysis.

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Research Organization:: SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 1628840

Journal Information:: eLife, Vol. 4; ISSN 2050-084X

Publisher:: eLife Sciences Publications, Ltd.Copyright Statement

Country of Publication:: United States

Language:: English

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