Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10795755 | Biochimica et Biophysica Acta (BBA) - Bioenergetics | 2014 | 7 Pages |
Abstract
The function of F1-ATPase relies critically on the intrinsic ability of its catalytic and noncatalytic subunits to interact with nucleotides. Therefore, the study of isolated subunits represents an opportunity to dissect elementary energetic contributions that drive the enzyme's rotary mechanism. In this study we have calorimetrically characterized the association of adenosine nucleotides to the isolated noncatalytic α-subunit. The resulting recognition behavior was compared with that previously reported for the isolated catalytic β-subunit (N.O. Pulido, G. Salcedo, G. Pérez-Hernández, C. José-Núñez, A. Velázquez-Campoy, E. GarcÃa-Hernández, Energetic effects of magnesium in the recognition of adenosine nucleotides by the F1-ATPase β subunit, Biochemistry 49 (2010) 5258-5268). The two subunits exhibit nucleotide-binding thermodynamic signatures similar to each other, characterized by enthalpically-driven affinities in the μM range. Nevertheless, contrary to the catalytic subunit that recognizes MgATP and MgADP with comparable strength, the noncatalytic subunit much prefers the triphosphate nucleotide. Besides, the α-subunit depends more on Mg(II) for stabilizing the interaction with ATP, while both subunits are rather metal-independent for ADP recognition. These binding behaviors are discussed in terms of the properties that the two subunits exhibit in the whole enzyme.
Keywords
ΔHbΔGBinding enthalpyMgCl2F1-ATPaseΔHMg(II)ITCTRISΔSEF1ADPnuclear magnetic resonance2-amino-2-(hydroxymethyl)propane-1,3-dioltf1adenosine 5′-triphosphateadenosine 5′-triphosphataseadenosine 5′-diphosphateATPMetal bindingEDTAEthylenediaminetetraacetic acidBinding energeticsATPaseNMRequilibrium dissociation constantEquilibrium binding constantIsothermal titration calorimetryMagnesium chloride
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Authors
Guillermo Salcedo, Patricia Cano-Sánchez, Marietta Tuena de Gómez-Puyou, Adrián Velázquez-Campoy, Enrique GarcÃa-Hernández,