The coupling ion in the methanoarchaeal ATP synthases: H(+) vs. Na(+) in the A(1)A(o) ATP synthase from the archaeon Methanosarcina mazei Gö1.
To establish a system to analyze ATP synthesis by the archaeal A(1)A(o) ATP synthase and to address the nature of the coupling ion, the operon encoding the A(1)A(o) ATP synthase from the mesophile Methanosarcina mazei Gö1 was cloned in an expression vector and it was expressed in the F(1)F(o) ATP synthase-negative mutant Escherichia coli DK8. Western blot analyses revealed that each of the subunits was produced, and the subunits assembled to a functional, membrane-embedded ATP synthase/ATPase. ATP hydrolysis was inhibited by dicyclohexylcarbodiimide but also by tributyltin, which turned out to be the most efficient inhibitor of the A(o) domain of A(1)A(o) ATP synthase known to date. ATP hydrolysis was not dependent on the Na(+) concentration of the medium, and inhibition of the enzyme by dicyclohexylcarbodiimide could not be relieved by Na(+). The enzyme present in the cytoplasmic membrane of E. coli catalyzed ATP synthesis driven by an artificial DeltapH but not by DeltapNa or DeltamuNa(+).
Pisa
Kim Y
KY
Molecular Microbiology & Bioenergetics, Institute of Molecular Biosciences, Johann Wolfgang Goethe Universität Frankfurt, Frankfurt, Germany.
Weidner
Claudia
C
Maischak
Heiko
H
Kavermann
Holger
H
Müller
Volker
V
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D003470:Culture Media; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D019848:Gene Expression Regulation, Archaeal; D006859:Hydrogen / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D017020:Methanosarcina / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism*
D019843:Archaeal Proteins; D003470:Culture Media; D006859:Hydrogen; D000255:Adenosine Triphosphate; D012964:Sodium; C113203:A1A0 ATPase, Methanosarcina mazei; D006180:Proton-Translocating ATPases