Metal Transfer within the Escherichia coli HypB–HypA Complex of Hydrogenase Accessory Proteins
| Autor: | Thanh T. Ngu, Deborah B. Zamble, Harini Kaluarachchi, Colin D. Douglas |
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| Rok vydání: | 2013 |
| Předmět: |
Spectrometry
Mass Electrospray Ionization Hydrogenase GTPase medicine.disease_cause Biochemistry Article Insert (molecular biology) GTP-binding protein regulators GTP-Binding Proteins Escherichia coli medicine Metal transfer DNA Primers Base Sequence biology Metal binding Chemistry Escherichia coli Proteins Intracellular Signaling Peptides and Proteins Active site Cell biology Metals biology.protein Carrier Proteins |
| Zdroj: | Biochemistry. 52:6030-6039 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi400812r |
| Popis: | The maturation of [NiFe]-hydrogenase in Escherichia coli is a complex process involving many steps and multiple accessory proteins. The two accessory proteins HypA and HypB interact with each other and are thought to cooperate to insert nickel into the active site of the hydrogenase-3 precursor protein. Both of these accessory proteins bind metal individually, but little is known about the metal-binding activities of the proteins once they assemble together into a functional complex. In this study, we investigate how complex formation modulates metal binding to the E. coli proteins HypA and HypB. This work lead to a re-evaluation of the HypA nickel affinity, revealing a KD on the order of 10(-8) M. HypA can efficiently remove nickel, but not zinc, from the metal-binding site in the GTPase domain of HypB, a process that is less efficient when complex formation between HypA and HypB is disrupted. Furthermore, nickel release from HypB to HypA is specifically accelerated when HypB is loaded with GDP, but not GTP. These results are consistent with the HypA-HypB complex serving as a transfer step in the relay of nickel from membrane transporter to its final destination in the hydrogenase active site and suggest that this complex contributes to the metal fidelity of this pathway. |
| Databáze: | OpenAIRE |
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