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

Abstract

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.

Authors:
 [1];  [2];  [3];  [1]
+ Show Author Affiliations
  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
  2. California Institute of Technology (CalTech), Pasadena, CA (United States). Division of Chemistry and Chemical Engineering
  3. 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
Publication Date:
Research Org.:
SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division
OSTI Identifier:
1628840
Grant/Contract Number:  
AC02-76SF00515
Resource Type:
Accepted Manuscript
Journal Name:
eLife
Additional Journal Information:
Journal Volume: 4; Journal ID: ISSN 2050-084X
Publisher:
eLife Sciences Publications, Ltd.
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Life Sciences & Biomedicine - Other Topics

Citation Formats

Spatzal, Thomas, Perez, Kathryn A., Howard, James B., and Rees, Douglas C. Catalysis-dependent selenium incorporation and migration in the nitrogenase active site iron-molybdenum cofactor. United States: N. p., 2015. Web. doi:10.7554/elife.11620.
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Spatzal, Thomas, Perez, Kathryn A., Howard, James B., & Rees, Douglas C. Catalysis-dependent selenium incorporation and migration in the nitrogenase active site iron-molybdenum cofactor. United States. https://doi.org/10.7554/elife.11620
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Spatzal, Thomas, Perez, Kathryn A., Howard, James B., and Rees, Douglas C. Wed . "Catalysis-dependent selenium incorporation and migration in the nitrogenase active site iron-molybdenum cofactor". United States. https://doi.org/10.7554/elife.11620. https://www.osti.gov/servlets/purl/1628840.
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@article{osti_1628840,
title = {Catalysis-dependent selenium incorporation and migration in the nitrogenase active site iron-molybdenum cofactor},
author = {Spatzal, Thomas and Perez, Kathryn A. and Howard, James B. and Rees, Douglas C.},
abstractNote = {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.},
doi = {10.7554/elife.11620},
journal = {eLife},
number = ,
volume = 4,
place = {United States},
year = {Wed Dec 16 00:00:00 EST 2015},
month = {Wed Dec 16 00:00:00 EST 2015}
}
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https://doi.org/10.7554/elife.11620
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    Works referencing / citing this record:

    The structure of vanadium nitrogenase reveals an unusual bridging ligand
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