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

Journal Article · Mon Jan 01 04:00:00 EST 2007 · Journal of Biological Inorganic Chemistry

DOI:https://doi.org/10.1007/s00775-007-0224-z· OSTI ID:944495

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

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.

• Research Organization: National Renewable Energy Laboratory (NREL), Golden, CO.
• Sponsoring Organization: USDOE
• DOE Contract Number: AC36-99GO10337
• OSTI ID: 944495
• Journal Information: Journal of Biological Inorganic Chemistry, Journal Name: Journal of Biological Inorganic Chemistry Journal Issue: 2007 Vol. 12
• Country of Publication: United States
• Language: English

Related Subjects

08 HYDROGEN
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
59 BASIC BIOLOGICAL SCIENCES
Chemical Technologies
ENZYMES
ESCHERICHIA COLI
HYDROGENASES
IN VITRO
PRECURSOR
PROTEINS
SUBSTRATES