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Catalysis by isolated β-subunits of the ATP Synthase/ATPase from Thermophilic bacillus PS3. Hydrolysis of Pyrophosphate

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Abstract

Although the capacity of isolated β-subunits of the ATP synthase/ATPase to perform catalysis has been extensively studied, the results have not conclusively shown that the subunits are catalytically active. Since soluble F1 of mitochondrial H+-ATPase can bind inorganic pyrophosphate (PPi) and synthesize PPi from medium phosphate, we examined if purified His-tagged β-subunits from Thermophilic bacillus PS3 can hydrolyze PPi. The difference spectra in the near UV CD of β-subunits with and without PPi show that PPi binds to the subunits. Other studies show that β-subunits hydrolyze [32P] PPi through a Mg2+-dependent process with an optimal pH of 8.3. Free Mg2+ is required for maximal hydrolytic rates. The Km for PPi is 75 μM and the Vmax is 800 pmol/min/mg. ATP is a weak inhibitor of the reaction, it diminishes the Vmax and increases the Km for PPi. Thus, isolated β-subunits are catalytically competent with PPi as substrate; apparently, the assembly of β-subunits into the ATPase complex changes substrate specificity, and leads to an increase in catalytic rates.

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Authors and Affiliations

  1. Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Ciudad Universitaria, Apartado Postal 70243, 04510, México, D. F., México

    Concepción José-Nuñez, Alfredo Torres-Larios, Armando Gómez-Puyou & Marietta Tuena de Gómez-Puyou

  2. Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de Mexico, México, D. F., México

    Leticia Ramírez-Silva & Guillermo Mendoza

  3. Facultad de Ciencias, Universidad Nacional Autónoma de México, Ciudad Universitaria, Apartado Postal 70243, 04510, México, D. F., México

    Guillermo Salcedo

Authors
  1. Concepción José-Nuñez
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  2. Alfredo Torres-Larios
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  3. Leticia Ramírez-Silva
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  4. Guillermo Mendoza
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  5. Guillermo Salcedo
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  6. Armando Gómez-Puyou
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  7. Marietta Tuena de Gómez-Puyou
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Correspondence to Marietta Tuena de Gómez-Puyou.

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José-Nuñez, C., Torres-Larios, A., Ramírez-Silva, L. et al. Catalysis by isolated β-subunits of the ATP Synthase/ATPase from Thermophilic bacillus PS3. Hydrolysis of Pyrophosphate. J Bioenerg Biomembr 40, 561–568 (2008). https://doi.org/10.1007/s10863-008-9192-4

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  • DOI: https://doi.org/10.1007/s10863-008-9192-4

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