GTP Hydrolysis by HypB is Essential for Nickel Insertion into Hydrogenases of Escherichia Coli
作者： Thomas Maier , Friedrich Lottspeich , August Bock
DOI: 10.1111/J.1432-1033.1995.0133I.X
关键词: Protein subunit 、 Escherichia coli 、 Biology 、 GTPase 、 Asparagine 、 GTP' 、 Mutant 、 Hydrogenase 、 Biochemistry 、 Enzyme kinetics 、 Cell biology 、 Molecular biology
摘要: The product of the hypB gene, which is required for maturation three [NiFe]hydrogenases Escherichia coli, a member GTPase family and exhibits low intrinsic activity. It was studied whether or not GTP hydrolysis by HypB coupled to nickel insertion into hydrogenases hydrogenases. Mutations were introduced gene at sites expected code amino acids involved in guanine-nucleotide binding. Lys117 G-motif 1, as well Asp241 4 substituted asparagine residues. purified mutant proteins showed strongly reduced, but still significant, In case [D241N]HypB, kcat/Km value lowered factor 85 specificity enzyme apparently lost, with other nucleoside triphosphates including XTP becoming compatible substrates. decrease activity even more pronounced [K117N]HypB. To assess functionality these vivo, wild-type chromosome E. coli replaced alleles. resulting strains BKN117 BDN241 affected hydrogen metabolism under fermentative conditions. did display hydrogenase due loss incorporation large subunit. exhibited reduction 44% only portion 3 subunit mature nickel-containing form. From results, it concluded that catalysed an integral process
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