Cloning and Functional Expression Analysis of the α Subunit of Mouse ATP Synthase

1993 ◽  
Vol 191 (1) ◽  
pp. 142-148 ◽  
Author(s):  
W.V. Yotov ◽  
R. Starnaud
Keyword(s):  
Atp Synthase ◽  
Expression Analysis ◽  
Functional Expression ◽  
Α Subunit

scholarly journals Targeting Mycobacterial F-ATP Synthase C-Terminal α Subunit Interaction Motif on Rotary Subunit γ

Antibiotics ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1456
Author(s):  
Amaravadhi Harikishore ◽  
Chui-Fann Wong ◽  
Priya Ragunathan ◽  
Dennis Litty ◽  
Volker Müller ◽  
...  
Keyword(s):  
Atp Synthase ◽  
Atp Hydrolysis ◽  
Atp Synthesis ◽  
Membrane Vesicles ◽  
Α Subunit ◽  
Rotational States ◽  
C Terminus ◽  
Inverted Membrane Vesicles ◽  
Hydrolysis Of ◽  
Micromolar Range

Mycobacteria regulate their energy (ATP) levels to sustain their survival even in stringent living conditions. Recent studies have shown that mycobacteria not only slow down their respiratory rate but also block ATP hydrolysis of the F-ATP synthase (α3:β3:γ:δ:ε:a:b:b’:c9) to maintain ATP homeostasis in situations not amenable for growth. The mycobacteria-specific α C-terminus (α533-545) has unraveled to be the major regulative of latent ATP hydrolysis. Its deletion stimulates ATPase activity while reducing ATP synthesis. In one of the six rotational states of F-ATP synthase, α533-545 has been visualized to dock deep into subunit γ, thereby blocking rotation of γ within the engine. The functional role(s) of this C-terminus in the other rotational states are not clarified yet and are being still pursued in structural studies. Based on the interaction pattern of the docked α533-545 region with subunit γ, we attempted to study the druggability of the α533-545 motif. In this direction, our computational work has led to the development of an eight-featured α533-545 peptide pharmacophore, followed by database screening, molecular docking, and pose selection, resulting in eleven hit molecules. ATP synthesis inhibition assays using recombinant ATP synthase as well as mycobacterial inverted membrane vesicles show that one of the hits, AlMF1, inhibited the mycobacterial F-ATP synthase in a micromolar range. The successful targeting of the α533-545-γ interaction motif demonstrates the potential to develop inhibitors targeting the α site to interrupt rotary coupling with ATP synthesis.

scholarly journals Structure of the ATP synthase from Mycobacterium smegmatis provides targets for treating tuberculosis

2021 ◽  
Vol 118 (47) ◽  
pp. e2111899118
Author(s):  
Martin G. Montgomery ◽  
Jessica Petri ◽  
Tobias E. Spikes ◽  
John E. Walker
Keyword(s):  
Atp Synthase ◽  
Mycobacterium Smegmatis ◽  
Atp Hydrolysis ◽  
Terminal Region ◽  
Catalytic Cycle ◽  
Α Subunit ◽  
Antitubercular Drugs ◽  
Safe Mechanism ◽  
Fail Safe ◽  
Α Subunits

The structure has been determined by electron cryomicroscopy of the adenosine triphosphate (ATP) synthase from Mycobacterium smegmatis. This analysis confirms features in a prior description of the structure of the enzyme, but it also describes other highly significant attributes not recognized before that are crucial for understanding the mechanism and regulation of the mycobacterial enzyme. First, we resolved not only the three main states in the catalytic cycle described before but also eight substates that portray structural and mechanistic changes occurring during a 360° catalytic cycle. Second, a mechanism of auto-inhibition of ATP hydrolysis involves not only the engagement of the C-terminal region of an α-subunit in a loop in the γ-subunit, as proposed before, but also a “fail-safe” mechanism involving the b′-subunit in the peripheral stalk that enhances engagement. A third unreported characteristic is that the fused bδ-subunit contains a duplicated domain in its N-terminal region where the two copies of the domain participate in similar modes of attachment of the two of three N-terminal regions of the α-subunits. The auto-inhibitory plus the associated “fail-safe” mechanisms and the modes of attachment of the α-subunits provide targets for development of innovative antitubercular drugs. The structure also provides support for an observation made in the bovine ATP synthase that the transmembrane proton-motive force that provides the energy to drive the rotary mechanism is delivered directly and tangentially to the rotor via a Grotthuss water chain in a polar L-shaped tunnel.

Ammonium transport by the colonic H+-K+-ATPase expressed in Xenopusoocytes

1999 ◽  
Vol 277 (2) ◽  
pp. C280-C287 ◽  
Author(s):  
Marc Cougnon ◽  
Patrice Bouyer ◽  
Frédéric Jaisser ◽  
Aleksander Edelman ◽  
Gabrielle Planelles
Keyword(s):  
Kinetic Analysis ◽  
Initial Rate ◽  
Functional Expression ◽  
Competitive Inhibitor ◽  
Ammonium Transport ◽  
Xenopus Laevis Oocytes ◽  
Intracellular Acidification ◽  
Extracellular Medium ◽  
Α Subunit ◽  
External Site

Functional expression of the rat colonic H+-K+-ATPase was obtained by coexpressing its catalytic α-subunit and the β1-subunit of the Na+-K+-ATPase in Xenopus laevis oocytes. We observed that, in oocytes expressing the rat colonic H+-K+-ATPase but not in control oocytes (expressing β1 alone), NH4Cl induced a decrease in86Rb uptake and the initial rate of intracellular acidification induced by extracellular NH4Cl was enhanced, consistent with [Formula: see text] influx via the colonic H+-K+-ATPase. In the absence of extracellular K+, only oocytes expressing the colonic H+-K+-ATPase were able to acidify an extracellular medium supplemented with NH4Cl. In the absence of extracellular K+ and in the presence of extracellular [Formula: see text], intracellular Na+ activity in oocytes expressing the colonic H+-K+-ATPase was lower than that in control oocytes. A kinetic analysis of86Rb uptake suggests that[Formula: see text] acts as a competitive inhibitor of the pump. Taken together, these results are consistent with[Formula: see text] competition for K+ on the external site of the colonic H+-K+-ATPase and with [Formula: see text] transport mediated by this pump.

scholarly journals Analysis of Sequence Determinants of F1Fo-ATP Synthase in the N-terminal Region of α Subunit for Binding of δ Subunit

2004 ◽  
Vol 279 (24) ◽  
pp. 25673-25679 ◽  
Author(s):  
Joachim Weber ◽  
Alma Muharemagic ◽  
Susan Wilke-Mounts ◽  
Alan E. Senior
Keyword(s):  
Atp Synthase ◽  
Terminal Region ◽  
Α Subunit ◽  
F1fo Atp Synthase

Molecular cloning and functional expression analysis of a cDNA for human hepassocin, a liver-specific protein with hepatocyte mitogenic activity

2001 ◽  
Vol 1520 (1) ◽  
pp. 45-53 ◽  
Author(s):  
Hiroshi Hara ◽  
Hiromitsu Yoshimura ◽  
Saeko Uchida ◽  
Yumiko Toyoda ◽  
Mari Aoki ◽  
...  
Keyword(s):  
Molecular Cloning ◽  
Expression Analysis ◽  
Functional Expression ◽  
Specific Protein ◽  
Mitogenic Activity ◽  
Liver Specific Protein

scholarly journals Regulation of the Nuclear Gene That Encodes the α-Subunit of the Mitochondrial F0F1-ATP Synthase Complex

1997 ◽  
Vol 272 (16) ◽  
pp. 10538-10542 ◽  
Author(s):  
Gail A. M. Breen ◽  
Elzora M. Jordan
Keyword(s):  
Atp Synthase ◽  
Nuclear Gene ◽  
Α Subunit ◽  
F0f1 Atp Synthase

Male-Biased Mutation Rates Revealed from Z and W Chromosome-Linked ATP Synthase α-Subunit (ATP5A1) Sequences in Birds

2000 ◽  
Vol 50 (5) ◽  
pp. 443-447 ◽  
Author(s):  
Ariane N. Carmichael ◽  
Anna-Karin Fridolfsson ◽  
Joy Halverson ◽  
Hans Ellegren
Keyword(s):  
Atp Synthase ◽  
Mutation Rates ◽  
W Chromosome ◽  
Α Subunit
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