Paradigm Shift for Radical S -Adenosyl- l -methionine Reactions: The Organometallic Intermediate Ω Is Central to Catalysis
Abstract
Radical S-adenosyl-L-methionine (SAM) enzymes comprise a vast superfamily catalyzing diverse reactions essential to all life through homolytic SAM cleavage to liberate the highly reactive 5'-deoxyadenosyl radical (5'-dAdo·). Our recent observation of a catalytically competent organometallic intermediate Ω that forms during reaction of the radical SAM (RS) enzyme pyruvate formate-lyase activating-enzyme (PFL-AE) was therefore quite surprising, and led to the question of its broad relevance in the superfamily. We now show that Ω in PFL-AE forms as an intermediate under a variety of mixing order conditions, suggesting it is central to catalysis in this enzyme. We further demonstrate that Ω forms in a suite of RS enzymes chosen to span the totality of superfamily reaction types, implicating Ω as essential in catalysis across the RS superfamily. Finally, EPR and electron nuclear double resonance spectroscopy establish that Ω involves an Fe-C5' bond between 5'-dAdo· and the [4Fe-4S] cluster. An analogous organometallic bond is found in the well-known adenosylcobalamin (coenzyme B12) cofactor used to initiate radical reactions via a 5'- dAdo· intermediate. Liberation of a reactive 5'-dAdo· intermediate via homolytic metal-carbon bond cleavage thus appears to be similar for Ω and coenzyme B12. However, coenzyme B12 is involved in enzymes catalyzing only amore »
- Authors:
-
- Department of Chemistry & Biochemistry, Montana State University, Bozeman, Montana 59717, United States
- Department of Chemistry, Northwestern University, Evanston, Illinois 60208, United States
- Institute of Microbiology, Eidgenössische Technische Hochschule Zürich, Vladimir-Prelog-Weg 4, Zürich 8093, Switzerland
- Cassia, LLC, 3030 Bunker Hill Street, Ste. 214, San Diego, California 92109, United States
- Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710, United States
- Publication Date:
- Research Org.:
- Montana State Univ., Bozeman, MT (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1459248
- Alternate Identifier(s):
- OSTI ID: 1508581
- Grant/Contract Number:
- SC0005404
- Resource Type:
- Published Article
- Journal Name:
- Journal of the American Chemical Society
- Additional Journal Information:
- Journal Name: Journal of the American Chemical Society Journal Volume: 140 Journal Issue: 28; Journal ID: ISSN 0002-7863
- Publisher:
- American Chemical Society
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
Citation Formats
Byer, Amanda S., Yang, Hao, McDaniel, Elizabeth C., Kathiresan, Venkatesan, Impano, Stella, Pagnier, Adrien, Watts, Hope, Denler, Carly, Vagstad, Anna L., Piel, Jörn, Duschene, Kaitlin S., Shepard, Eric M., Shields, Thomas P., Scott, Lincoln G., Lilla, Edward A., Yokoyama, Kenichi, Broderick, William E., Hoffman, Brian M., and Broderick, Joan B. Paradigm Shift for Radical S -Adenosyl- l -methionine Reactions: The Organometallic Intermediate Ω Is Central to Catalysis. United States: N. p., 2018.
Web. doi:10.1021/jacs.8b04061.
Byer, Amanda S., Yang, Hao, McDaniel, Elizabeth C., Kathiresan, Venkatesan, Impano, Stella, Pagnier, Adrien, Watts, Hope, Denler, Carly, Vagstad, Anna L., Piel, Jörn, Duschene, Kaitlin S., Shepard, Eric M., Shields, Thomas P., Scott, Lincoln G., Lilla, Edward A., Yokoyama, Kenichi, Broderick, William E., Hoffman, Brian M., & Broderick, Joan B. Paradigm Shift for Radical S -Adenosyl- l -methionine Reactions: The Organometallic Intermediate Ω Is Central to Catalysis. United States. https://doi.org/10.1021/jacs.8b04061
Byer, Amanda S., Yang, Hao, McDaniel, Elizabeth C., Kathiresan, Venkatesan, Impano, Stella, Pagnier, Adrien, Watts, Hope, Denler, Carly, Vagstad, Anna L., Piel, Jörn, Duschene, Kaitlin S., Shepard, Eric M., Shields, Thomas P., Scott, Lincoln G., Lilla, Edward A., Yokoyama, Kenichi, Broderick, William E., Hoffman, Brian M., and Broderick, Joan B. Thu .
"Paradigm Shift for Radical S -Adenosyl- l -methionine Reactions: The Organometallic Intermediate Ω Is Central to Catalysis". United States. https://doi.org/10.1021/jacs.8b04061.
@article{osti_1459248,
title = {Paradigm Shift for Radical S -Adenosyl- l -methionine Reactions: The Organometallic Intermediate Ω Is Central to Catalysis},
author = {Byer, Amanda S. and Yang, Hao and McDaniel, Elizabeth C. and Kathiresan, Venkatesan and Impano, Stella and Pagnier, Adrien and Watts, Hope and Denler, Carly and Vagstad, Anna L. and Piel, Jörn and Duschene, Kaitlin S. and Shepard, Eric M. and Shields, Thomas P. and Scott, Lincoln G. and Lilla, Edward A. and Yokoyama, Kenichi and Broderick, William E. and Hoffman, Brian M. and Broderick, Joan B.},
abstractNote = {Radical S-adenosyl-L-methionine (SAM) enzymes comprise a vast superfamily catalyzing diverse reactions essential to all life through homolytic SAM cleavage to liberate the highly reactive 5'-deoxyadenosyl radical (5'-dAdo·). Our recent observation of a catalytically competent organometallic intermediate Ω that forms during reaction of the radical SAM (RS) enzyme pyruvate formate-lyase activating-enzyme (PFL-AE) was therefore quite surprising, and led to the question of its broad relevance in the superfamily. We now show that Ω in PFL-AE forms as an intermediate under a variety of mixing order conditions, suggesting it is central to catalysis in this enzyme. We further demonstrate that Ω forms in a suite of RS enzymes chosen to span the totality of superfamily reaction types, implicating Ω as essential in catalysis across the RS superfamily. Finally, EPR and electron nuclear double resonance spectroscopy establish that Ω involves an Fe-C5' bond between 5'-dAdo· and the [4Fe-4S] cluster. An analogous organometallic bond is found in the well-known adenosylcobalamin (coenzyme B12) cofactor used to initiate radical reactions via a 5'- dAdo· intermediate. Liberation of a reactive 5'-dAdo· intermediate via homolytic metal-carbon bond cleavage thus appears to be similar for Ω and coenzyme B12. However, coenzyme B12 is involved in enzymes catalyzing only a small number (~12) of distinct reactions, whereas the RS superfamily has more than 100 000 distinct sequences and over 80 reaction types characterized to date. The appearance of Ω across the RS superfamily therefore dramatically enlarges the sphere of bio-organometallic chemistry in Nature.},
doi = {10.1021/jacs.8b04061},
journal = {Journal of the American Chemical Society},
number = 28,
volume = 140,
place = {United States},
year = {2018},
month = {6}
}
https://doi.org/10.1021/jacs.8b04061
Web of Science
Figures / Tables:

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