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The fully-active and structurally-stable form of the mitochondrial ATP synthase of Polytomella sp. is dimeric

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Abstract

Mitochondrial F1FO-ATP synthase of chlorophycean algae is a stable dimeric complex of 1,600 kDa. It lacks the classic subunits that constitute the peripheral stator-stalk and the orthodox polypeptides involved in the dimerization of the complex. Instead, it contains nine polypeptides of unknown evolutionary origin named ASA1 to ASA9. The isolated enzyme exhibited a very low ATPase activity (0.03 Units/mg), that increased upon heat treatment, due to the release of the F1 sector. Oligomycin was found to stabilize the dimeric structure of the enzyme, providing partial resistance to heat dissociation. Incubation in the presence of low concentrations of several non-ionic detergents increased the oligomycin-sensitive ATPase activity up to 7.0–9.0 Units/mg. Incubation with 3% (w/v) taurodeoxycholate monomerized the enzyme. The monomeric form of the enzyme exhibited diminished activity in the presence of detergents and diminished oligomycin sensitivity. Cross-linking experiments carried out with the dimeric and monomeric forms of the ATP synthase suggested the participation of the ASA6 subunit in the dimerization of the enzyme. The dimeric enzyme was more resistant to heat treatment, high hydrostatic pressures, and protease digestion than the monomeric enzyme, which was readily disrupted by these treatments. We conclude that the fully-active algal mitochondrial ATP synthase is a stable catalytically active dimer; the monomeric form is less active and less stable. Monomer-monomer interactions could be mediated by the membrane-bound subunits ASA6 and ASA9, and may be further stabilized by other polypeptides such as ASA1 and ASA5.

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Authors and Affiliations

  1. Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, México D.F., Mexico

    Alexa Villavicencio-Queijeiro, Miriam Vázquez-Acevedo, Araceli Cano-Estrada, Mariel Zarco-Zavala, Marietta Tuena de Gómez & Diego González-Halphen

  2. Instituto de Bioquímica Médica, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil

    Julio A. Mignaco, Monica M. Freire, Helena M. Scofano & Debora Foguel

  3. Genetics of Microorganisms, Institute of Plant Biology, University of Liège, Liège, Belgium

    Pierre Cardol & Claire Remacle

  4. Departamento de Genética Molecular, Instituto de Fisiología Celular, UNAM, Apartado Postal 70-600, Delegación Coyoacán, 04510, México D.F., México

    Diego González-Halphen

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  1. Alexa Villavicencio-Queijeiro
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  2. Miriam Vázquez-Acevedo
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  3. Araceli Cano-Estrada
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  4. Mariel Zarco-Zavala
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  5. Marietta Tuena de Gómez
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  6. Julio A. Mignaco
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  7. Monica M. Freire
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  8. Helena M. Scofano
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  9. Debora Foguel
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  10. Pierre Cardol
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  11. Claire Remacle
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  12. Diego González-Halphen
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Correspondence to Diego González-Halphen.

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Alexa Villavicencio-Queijeiro and Miriam Vázquez-Acevedo have contributed equally to this work.

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Villavicencio-Queijeiro, A., Vázquez-Acevedo, M., Cano-Estrada, A. et al. The fully-active and structurally-stable form of the mitochondrial ATP synthase of Polytomella sp. is dimeric. J Bioenerg Biomembr 41, 1–13 (2009). https://doi.org/10.1007/s10863-009-9203-0

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