Structural characterization of subunit b of the bacterial F1FO ATP synthase and the critical roles of conserved P-loop residues of the archaea A1AO ATP synthase subunit A

Adenosine 5’-triphosphate (ATP) synthesis by oxidative phosphorylation or photophosphorylation is a multistep membrane-located process that provides the bulk of cellular energy in eukaryotes and many prokaryotes. The majority of ATP synthesis is accomplished by the enzyme ATP synthase (EC 3.6.1.34)...

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Main Author: Ragunathan Priya
Other Authors: Gerhard Gruber
Format: Theses and Dissertations
Language:English
Published: 2011
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Online Access:http://hdl.handle.net/10356/44857
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-448572023-02-28T18:32:58Z Structural characterization of subunit b of the bacterial F1FO ATP synthase and the critical roles of conserved P-loop residues of the archaea A1AO ATP synthase subunit A Ragunathan Priya Gerhard Gruber School of Biological Sciences DRNTU::Science::Biological sciences::Microbiology::Bacteria Adenosine 5’-triphosphate (ATP) synthesis by oxidative phosphorylation or photophosphorylation is a multistep membrane-located process that provides the bulk of cellular energy in eukaryotes and many prokaryotes. The majority of ATP synthesis is accomplished by the enzyme ATP synthase (EC 3.6.1.34) also called as F1FO ATP synthase, which, in its simplest form, as in bacteria, is composed of eight different subunits. This multi-subunit complex is divided into the F1 headpiece, α3:β3, attached by a central and a peripheral stalk to a membrane-embedded ion-translocating part known as FO. The central stalk is made up of γ and ε and the peripheral stalk consists of b and δ. As the central stalk is driven by the passage of protons through a pore formed by the c and a subunits of FO, the rotation of γ within α3β3 of F1 causes conformational changes in the catalytic nucleotide binding sites in β-subunits, resulting in ATP synthesis and release. One main function of the b2δ stator is to hold the α3β3 hexamer against the rotational torque caused by the rotation of the central stalk. The b subunit dimer in the peripheral stator stalk of Escherichia coli ATP synthase is essential for enzyme assembly and the rotational catalytic mechanism. In order to get the overall structural details, subunit b, which makes the peripheral stalk of F1FO ATP synthase, was expressed in E. coli cells as a soluble protein and purified to homogeneity. The first low resolution structure of the soluble domain of subunit b (b22-156) was determined from solution X-ray scattering data. The protein has a boomerang-like shape with a total length of 16.2 ± 0.3 nm. Independent to the protein concentration used, b22-156 is dimeric in solution. Circular dichroism (CD) spectroscopy revealed that b22-156 comprises of 77% α-helix. Doctor of Philosophy (SBS) 2011-06-06T06:31:46Z 2011-06-06T06:31:46Z 2011 2011 Thesis http://hdl.handle.net/10356/44857 en 150 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science::Biological sciences::Microbiology::Bacteria
spellingShingle DRNTU::Science::Biological sciences::Microbiology::Bacteria
Ragunathan Priya
Structural characterization of subunit b of the bacterial F1FO ATP synthase and the critical roles of conserved P-loop residues of the archaea A1AO ATP synthase subunit A
description Adenosine 5’-triphosphate (ATP) synthesis by oxidative phosphorylation or photophosphorylation is a multistep membrane-located process that provides the bulk of cellular energy in eukaryotes and many prokaryotes. The majority of ATP synthesis is accomplished by the enzyme ATP synthase (EC 3.6.1.34) also called as F1FO ATP synthase, which, in its simplest form, as in bacteria, is composed of eight different subunits. This multi-subunit complex is divided into the F1 headpiece, α3:β3, attached by a central and a peripheral stalk to a membrane-embedded ion-translocating part known as FO. The central stalk is made up of γ and ε and the peripheral stalk consists of b and δ. As the central stalk is driven by the passage of protons through a pore formed by the c and a subunits of FO, the rotation of γ within α3β3 of F1 causes conformational changes in the catalytic nucleotide binding sites in β-subunits, resulting in ATP synthesis and release. One main function of the b2δ stator is to hold the α3β3 hexamer against the rotational torque caused by the rotation of the central stalk. The b subunit dimer in the peripheral stator stalk of Escherichia coli ATP synthase is essential for enzyme assembly and the rotational catalytic mechanism. In order to get the overall structural details, subunit b, which makes the peripheral stalk of F1FO ATP synthase, was expressed in E. coli cells as a soluble protein and purified to homogeneity. The first low resolution structure of the soluble domain of subunit b (b22-156) was determined from solution X-ray scattering data. The protein has a boomerang-like shape with a total length of 16.2 ± 0.3 nm. Independent to the protein concentration used, b22-156 is dimeric in solution. Circular dichroism (CD) spectroscopy revealed that b22-156 comprises of 77% α-helix.
author2 Gerhard Gruber
author_facet Gerhard Gruber
Ragunathan Priya
format Theses and Dissertations
author Ragunathan Priya
author_sort Ragunathan Priya
title Structural characterization of subunit b of the bacterial F1FO ATP synthase and the critical roles of conserved P-loop residues of the archaea A1AO ATP synthase subunit A
title_short Structural characterization of subunit b of the bacterial F1FO ATP synthase and the critical roles of conserved P-loop residues of the archaea A1AO ATP synthase subunit A
title_full Structural characterization of subunit b of the bacterial F1FO ATP synthase and the critical roles of conserved P-loop residues of the archaea A1AO ATP synthase subunit A
title_fullStr Structural characterization of subunit b of the bacterial F1FO ATP synthase and the critical roles of conserved P-loop residues of the archaea A1AO ATP synthase subunit A
title_full_unstemmed Structural characterization of subunit b of the bacterial F1FO ATP synthase and the critical roles of conserved P-loop residues of the archaea A1AO ATP synthase subunit A
title_sort structural characterization of subunit b of the bacterial f1fo atp synthase and the critical roles of conserved p-loop residues of the archaea a1ao atp synthase subunit a
publishDate 2011
url http://hdl.handle.net/10356/44857
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