scholarly journals The Escherichia coli PriA Helicase–Double-Stranded DNA Complex: Location of the Strong DNA-Binding Subsite on the Helicase Domain of the Protein and the Affinity Control by the Two Nucleotide-Binding Sites of the Enzyme

2010 ◽  
Vol 402 (2) ◽  
pp. 344-362 ◽  
Author(s):  
Michal R. Szymanski ◽  
Maria J. Jezewska ◽  
Wlodzimierz Bujalowski
Keyword(s):  
Escherichia Coli ◽  
Dna Binding ◽  
Binding Sites ◽  
Nucleotide Binding ◽  
Helicase Domain ◽  
Nucleotide Binding Sites ◽  
Double Stranded Dna ◽  
Dna Complex

scholarly journals Observation of Asymmetry amongst Nucleotide Binding Sites of F1-ATPase of Escherichia coli by31P NMR Spectroscopy

2011 ◽  
Vol 32 (2) ◽  
pp. 531-535 ◽  
Author(s):  
Nam-Kung Jun ◽  
Joon-Hyung Sohn ◽  
Byung-Il Yeh ◽  
Jong-Whan Choi ◽  
Hyun-Won Kim
Keyword(s):  
Escherichia Coli ◽  
Nmr Spectroscopy ◽  
Binding Sites ◽  
Nucleotide Binding ◽  
F1 Atpase ◽  
Nucleotide Binding Sites

scholarly journals Energy transfer between the nicotinamide nucleotide transhydrogenase and ATP synthase of Escherichia coli

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Simone Sandra Graf ◽  
Sangjin Hong ◽  
Philipp Müller ◽  
Robert Gennis ◽  
Christoph von Ballmoos
Keyword(s):  
Escherichia Coli ◽  
Atp Synthase ◽  
Binding Sites ◽  
Proton Transport ◽  
Hydride Transfer ◽  
Nucleotide Binding ◽  
Coupling Mechanism ◽  
Long Distance ◽  
Nicotinamide Nucleotide Transhydrogenase ◽  
Nucleotide Binding Sites

AbstractMembrane bound nicotinamide nucleotide transhydrogenase (TH) catalyses the hydride transfer from NADH to NADP+. Under physiological conditions, this reaction is endergonic and must be energized by the pmf, coupled to transmembrane proton transport. Recent structures of transhydrogenase holoenzymes suggest new mechanistic details, how the long-distance coupling between hydride transfer in the peripheral nucleotide binding sites and the membrane-localized proton transfer occurs that now must be tested experimentally. Here, we provide protocols for the efficient expression and purification of the Escherichia coli transhydrogenase and its reconstitution into liposomes, alone or together with the Escherichia coli F1F0 ATP synthase. We show that E. coli transhydrogenase is a reversible enzyme that can also work as a NADPH-driven proton pump. In liposomes containing both enzymes, NADPH driven H+-transport by TH is sufficient to instantly fuel ATP synthesis, which adds TH to the pool of pmf generating enzymes. If the same liposomes are energized with ATP, NADPH production by TH is stimulated > sixfold both by a pH gradient or a membrane potential. The presented protocols and results reinforce the tight coupling between hydride transfer in the peripheral nucleotide binding sites and transmembrane proton transport and provide powerful tools to investigate their coupling mechanism.

Site-directed mutagenesis of tyrosine residues at nicotinamide nucleotide binding sites of Escherichia coli transhydrogenase

Biochemistry ◽  
1993 ◽  
Vol 32 (48) ◽  
pp. 13237-13244 ◽  
Author(s):  
Torbjoern Olausson ◽  
Thomas Hultman ◽  
Erland Holmberg ◽  
Jan Rydstroem ◽  
Suhail Ahmad ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Binding Sites ◽  
Nucleotide Binding ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Tyrosine Residues ◽  
Nucleotide Binding Sites
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