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Title: Equilibrium and ultrafast kinetic studies manipulating electron transfer: A short-lived flavin semiquinone is not sufficient for electron bifurcation

Abstract

Flavin-based electron transfer bifurcation is emerging as a fundamental and powerful mechanism for conservation and deployment of electrochemical energy in enzymatic systems. In this process, a pair of electrons is acquired at intermediate reduction potential (i.e. intermediate reducing power) and each electron is passed to a different acceptor, one with lower and the other with higher reducing power, leading to 'bifurcation'. It is believed that a strongly reducing semiquinone species is essential for this process, and it is expected that this species should be kinetically short-lived. We now demonstrate that presence of a short-lived anionic flavin semiquinone (ASQ) is not sufficient to infer existence of bifurcating activity, although such a species may be necessary for the process. We have used transient absorption spectroscopy to compare the rates and mechanisms of decay of ASQ generated photochemically in bifurcating NADH-dependent ferredoxin-NADP+ oxidoreductase and the non-bifurcating flavoproteins nitroreductase, NADH oxidase and flavodoxin. We found that different mechanisms dominate ASQ decay in the different protein environments, producing lifetimes ranging over two orders of magnitude. Capacity for electron transfer among redox cofactors vs. charge recombination with nearby donors can explain the range of ASQ lifetimes we observe. In conclusion, our results support a model whereinmore » efficient electron propagation can explain the short lifetime of the ASQ of bifurcating NADH-dependent ferredoxin-NADP+ oxidoreductase I, and can be an indication of capacity for electron bifurcation.« less

Authors:
; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1769200
Alternate Identifier(s):
OSTI ID: 1395083
Report Number(s):
NREL/JA-2700-68454
Journal ID: ISSN 0021-9258; S0021925820342228; PII: S0021925820342228
Grant/Contract Number:  
SC0012518; AC36–08-GO28308; AC36-08GO28308
Resource Type:
Published Article
Journal Name:
Journal of Biological Chemistry
Additional Journal Information:
Journal Name: Journal of Biological Chemistry Journal Volume: 292 Journal Issue: 34; Journal ID: ISSN 0021-9258
Publisher:
Elsevier
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; electron transfer; energetics; flavin; flavoprotein; fluorescence; electron bifurcation; transient absorption spectrocopy

Citation Formats

Hoben, John P., Lubner, Carolyn E., Ratzloff, Michael W., Schut, Gerrit J., Nguyen, Diep M. N., Hempel, Karl W., Adams, Michael W. W., King, Paul W., and Miller, Anne-Frances. Equilibrium and ultrafast kinetic studies manipulating electron transfer: A short-lived flavin semiquinone is not sufficient for electron bifurcation. United States: N. p., 2017. Web. doi:10.1074/jbc.M117.794214.
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Hoben, John P., Lubner, Carolyn E., Ratzloff, Michael W., Schut, Gerrit J., Nguyen, Diep M. N., Hempel, Karl W., Adams, Michael W. W., King, Paul W., & Miller, Anne-Frances. Equilibrium and ultrafast kinetic studies manipulating electron transfer: A short-lived flavin semiquinone is not sufficient for electron bifurcation. United States. https://doi.org/10.1074/jbc.M117.794214
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Hoben, John P., Lubner, Carolyn E., Ratzloff, Michael W., Schut, Gerrit J., Nguyen, Diep M. N., Hempel, Karl W., Adams, Michael W. W., King, Paul W., and Miller, Anne-Frances. Tue . "Equilibrium and ultrafast kinetic studies manipulating electron transfer: A short-lived flavin semiquinone is not sufficient for electron bifurcation". United States. https://doi.org/10.1074/jbc.M117.794214.
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@article{osti_1769200,
title = {Equilibrium and ultrafast kinetic studies manipulating electron transfer: A short-lived flavin semiquinone is not sufficient for electron bifurcation},
author = {Hoben, John P. and Lubner, Carolyn E. and Ratzloff, Michael W. and Schut, Gerrit J. and Nguyen, Diep M. N. and Hempel, Karl W. and Adams, Michael W. W. and King, Paul W. and Miller, Anne-Frances},
abstractNote = {Flavin-based electron transfer bifurcation is emerging as a fundamental and powerful mechanism for conservation and deployment of electrochemical energy in enzymatic systems. In this process, a pair of electrons is acquired at intermediate reduction potential (i.e. intermediate reducing power) and each electron is passed to a different acceptor, one with lower and the other with higher reducing power, leading to 'bifurcation'. It is believed that a strongly reducing semiquinone species is essential for this process, and it is expected that this species should be kinetically short-lived. We now demonstrate that presence of a short-lived anionic flavin semiquinone (ASQ) is not sufficient to infer existence of bifurcating activity, although such a species may be necessary for the process. We have used transient absorption spectroscopy to compare the rates and mechanisms of decay of ASQ generated photochemically in bifurcating NADH-dependent ferredoxin-NADP+ oxidoreductase and the non-bifurcating flavoproteins nitroreductase, NADH oxidase and flavodoxin. We found that different mechanisms dominate ASQ decay in the different protein environments, producing lifetimes ranging over two orders of magnitude. Capacity for electron transfer among redox cofactors vs. charge recombination with nearby donors can explain the range of ASQ lifetimes we observe. In conclusion, our results support a model wherein efficient electron propagation can explain the short lifetime of the ASQ of bifurcating NADH-dependent ferredoxin-NADP+ oxidoreductase I, and can be an indication of capacity for electron bifurcation.},
doi = {10.1074/jbc.M117.794214},
journal = {Journal of Biological Chemistry},
number = 34,
volume = 292,
place = {United States},
year = {Tue Aug 01 00:00:00 EDT 2017},
month = {Tue Aug 01 00:00:00 EDT 2017}
}
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https://doi.org/10.1074/jbc.M117.794214
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    Material descriptors for photocatalyst/catalyst design
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