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Title: In Vitro Activation of [FeFe] Hydrogenase: New Insights into Hydrogenase Maturation

Abstract

The in vitro activation of the [FeFe] hydrogenase is accomplished by combining Escherichia coli cell extracts containing the heterologously expressed inactive HydA with extracts in which hydrogenase-specific maturation proteins HydE, HydF, and HydG are expressed in concert. Interestingly, the process of HydA activation occurs rapidly and in the absence of potential substrates, which suggests that the hydrogenase accessory proteins synthesize an H-cluster precursor that can be quickly transferred to the hydrogenase enzyme to affect activation. HydA activity is observed to be dependent on the protein fraction containing all three accessory proteins expressed in concert and cannot be accomplished with addition of heat-treated extract or extract filtrate, suggesting that the activation of the hydrogenase structural protein is mediated by interaction with the accessory assembly protein(s). These results represent the first important step in understanding the process of H-cluster assembly and provide significant insights into hydrogenase maturation.

Authors:
; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
944495
DOE Contract Number:
AC36-99-GO10337
Resource Type:
Journal Article
Resource Relation:
Journal Name: Journal of Biological Inorganic Chemistry; Journal Volume: 12; Journal Issue: 2007
Country of Publication:
United States
Language:
English
Subject:
08 HYDROGEN; 37 INORGANIC, ORGANIC, PHYSICAL AND ANALYTICAL CHEMISTRY; 59 BASIC BIOLOGICAL SCIENCES; ENZYMES; ESCHERICHIA COLI; HYDROGENASES; IN VITRO; PRECURSOR; PROTEINS; SUBSTRATES; Chemical Technologies

Citation Formats

McGlynn, S. E., Ruebush, S. S., Naumov, A., Nagy, L. E., Dubini, A., King, P. W., Broderick, J. B., Posewitz, M. C., and Peters, J. W. In Vitro Activation of [FeFe] Hydrogenase: New Insights into Hydrogenase Maturation. United States: N. p., 2007. Web. doi:10.1007/s00775-007-0224-z.
McGlynn, S. E., Ruebush, S. S., Naumov, A., Nagy, L. E., Dubini, A., King, P. W., Broderick, J. B., Posewitz, M. C., & Peters, J. W. In Vitro Activation of [FeFe] Hydrogenase: New Insights into Hydrogenase Maturation. United States. doi:10.1007/s00775-007-0224-z.
McGlynn, S. E., Ruebush, S. S., Naumov, A., Nagy, L. E., Dubini, A., King, P. W., Broderick, J. B., Posewitz, M. C., and Peters, J. W. Mon . "In Vitro Activation of [FeFe] Hydrogenase: New Insights into Hydrogenase Maturation". United States. doi:10.1007/s00775-007-0224-z.
@article{osti_944495,
title = {In Vitro Activation of [FeFe] Hydrogenase: New Insights into Hydrogenase Maturation},
author = {McGlynn, S. E. and Ruebush, S. S. and Naumov, A. and Nagy, L. E. and Dubini, A. and King, P. W. and Broderick, J. B. and Posewitz, M. C. and Peters, J. W.},
abstractNote = {The in vitro activation of the [FeFe] hydrogenase is accomplished by combining Escherichia coli cell extracts containing the heterologously expressed inactive HydA with extracts in which hydrogenase-specific maturation proteins HydE, HydF, and HydG are expressed in concert. Interestingly, the process of HydA activation occurs rapidly and in the absence of potential substrates, which suggests that the hydrogenase accessory proteins synthesize an H-cluster precursor that can be quickly transferred to the hydrogenase enzyme to affect activation. HydA activity is observed to be dependent on the protein fraction containing all three accessory proteins expressed in concert and cannot be accomplished with addition of heat-treated extract or extract filtrate, suggesting that the activation of the hydrogenase structural protein is mediated by interaction with the accessory assembly protein(s). These results represent the first important step in understanding the process of H-cluster assembly and provide significant insights into hydrogenase maturation.},
doi = {10.1007/s00775-007-0224-z},
journal = {Journal of Biological Inorganic Chemistry},
number = 2007,
volume = 12,
place = {United States},
year = {Mon Jan 01 00:00:00 EST 2007},
month = {Mon Jan 01 00:00:00 EST 2007}
}