The fully-active and structurally-stable form of the mitochondrial ATP synthase of Polytomella sp. is dimeric.
Mitochondrial F(1)F(O)-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 F(1) 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.
Villavicencio-Queijeiro
Alexa
A
Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, México DF, Mexico.
Vázquez-Acevedo
Miriam
M
Cano-Estrada
Araceli
A
Zarco-Zavala
Mariel
M
Tuena de Gómez
Marietta
M
Mignaco
Julio A
JA
Freire
Monica M
MM
Scofano
Helena M
HM
Foguel
Debora
D
Cardol
Pierre
P
Remacle
Claire
C
González-Halphen
Diego
D
D000460:Chlorophyta / Q000201:enzymology*; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D025261:Mitochondrial Proton-Translocating ATPases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D011487:Protein Conformation; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism
D021122:Protein Subunits; D025261:Mitochondrial Proton-Translocating ATPases