Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/82139
Title: The stimulating role of subunit F in ATPase activity inside the A1-complex of the Methanosarcina mazei Gö1 A1AO ATP synthase
Authors: Singh, Dhirendra
Sielaff, Hendrik
Sundararaman, Lavanya
Bhushan, Shashi
Grüber, Gerhard
Keywords: A1AO ATP synthase
Subunit F
Issue Date: 2016
Source: Singh, D., Sielaff, H., Sundararaman, L., Bhushan, S., & Grüber, G. (2016). The stimulating role of subunit F in ATPase activity inside the A1-complex of the Methanosarcina mazei Gö1 A1AO ATP synthase. Biochimica et Biophysica Acta (BBA) - Bioenergetics, 1857(2), 177-187.
Series/Report no.: Biochimica et Biophysica Acta (BBA) - Bioenergetics
Abstract: A1AO ATP synthases couple ion-transport of the AO sector and ATP synthesis/hydrolysis of the A3B3-headpiece via their stalk subunits D and F. Here, we produced and purified stable A3B3D- and A3B3DF-complexes of the Methanosarcina mazei Gö1 A-ATP synthase as confirmed by electron microscopy. Enzymatic studies with these complexes showed that the M. mazei Gö1 A-ATP synthase subunit F is an ATPase activating subunit. The maximum ATP hydrolysis rates (Vmax) of A3B3D and A3B3DF were determined by substrate-dependent ATP hydrolysis experiments resulting in a Vmax of 7.9 s− 1 and 30.4 s− 1, respectively, while the KM is the same for both. Deletions of the N- or C-termini of subunit F abolished the effect of ATP hydrolysis activation. We generated subunit F mutant proteins with single amino acid substitutions and demonstrated that the subunit F residues S84 and R88 are important in stimulating ATP hydrolysis. Hybrid formation of the A3B3D-complex with subunit F of the related eukaryotic V-ATPase of Saccharomyces cerevisiae or subunit ε of the F-ATP synthase from Mycobacterium tuberculosis showed that subunit F of the archaea and eukaryotic enzymes are important in ATP hydrolysis.
URI: https://hdl.handle.net/10356/82139
http://hdl.handle.net/10220/41122
ISSN: 0005-2728
DOI: 10.1016/j.bbabio.2015.12.003
Rights: © 2016 Elsevier B.V. This is the author created version of a work that has been peer reviewed and accepted for publication by Biochimica et Biophysica Acta (BBA) - Bioenergetics, Elsevier B.V. It incorporates referee’s comments but changes resulting from the publishing process, such as copyediting, structural formatting, may not be reflected in this document. The published version is available at: [http://dx.doi.org/10.1016/j.bbabio.2015.12.003].
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SBS Journal Articles

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