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Please use this identifier to cite or link to this item: http://hdl.handle.net/1807/25441

Title: The Role of Dimerization by Escherichia coli HypB in Hydrogenase Biosynthesis
Authors: Cai, Fang
Advisor: Zamble, Deborah
Department: Chemistry
Keywords: Escherichia coli
HypB
Dimerization
Hydrogenase
Issue Date: 15-Dec-2010
Abstract: Nickel insertion into the [NiFe]-hydrogenase requires the accessory protein HypB, which is a GTPase. The GTPase domain of Escherichia coli (E. coli) HypB undergoes dimerization in the presence of GTP. To determine the role of HypB dimerization in hydrogenase biosynthesis, a double mutation L242A/L246A was introduced into full-length E. coli HypB, and the protein was expressed and characterized both in vitro and in vivo. Gel filtration experiments demonstrated that L242A/L246A HypB was monomeric as expected. The inability of L242A/L246A HypB to dimerize does not abolish its GTPase activity and the monomeric L242A/L246A HypB has a similar Ni(II)-binding behavior as that of wild type HypB. Upon the expression of L242A/L246A HypB in vivo the hydrogenase activity is approximately half of the activity of the wild-type control. These experimental results suggest that dimerization of HypB does have a, but not critical, role in hydrogenase biosynthesis.
URI: http://hdl.handle.net/1807/25441
Appears in Collections:Master
Department of Chemistry - Master theses

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