title: Oscillating Electric Field Measures the Rotation Rate in a Native Rotary Enzyme.
abstract: Rotary enzymes are complex, highly challenging biomolecular machines whose biochemical working mechanism involves intersubunit rotation. The true intrinsic rate of rotation of any rotary enzyme is not known in a native, unmodified state. Here we use the effect of an oscillating electric (AC) field on the biochemical activity of a rotary enzyme, the vacuolar proton-ATPase (V-ATPase), to directly measure its mean rate of rotation in its native membrane environment, without any genetic, chemical or mechanical modification of the enzyme, for the first time. The results suggest that a transmembrane AC field is able to synchronise the steps of ion-pumping in individual enzymes via a hold-and-release mechanism, which opens up the possibility of biotechnological exploitation. Our approach is likely to work for other transmembrane ion-transporting assemblies, not only rotary enzymes, to determine intrinsic in situ rates of ion pumping.
authors.affiliation: Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Temesvári krt. 62, 6726, Szeged, Hungary.
authors.forename: Csilla-Maria
authors.initials: CM
authors.lastname: Ferencz
authors.affiliation: Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Temesvári krt. 62, 6726, Szeged, Hungary.
authors.forename: Pál
authors.initials: P
authors.lastname: Petrovszki
authors.affiliation: Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Temesvári krt. 62, 6726, Szeged, Hungary.
authors.forename: András
authors.initials: A
authors.lastname: Dér
authors.affiliation: Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Temesvári krt. 62, 6726, Szeged, Hungary.
authors.forename: Krisztina
authors.initials: K
authors.lastname: Sebők-Nagy
authors.affiliation: Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Temesvári krt. 62, 6726, Szeged, Hungary.
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Kóta
authors.affiliation: Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Temesvári krt. 62, 6726, Szeged, Hungary.
authors.forename: Tibor
authors.initials: T
authors.lastname: Páli
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D011522:Protons; D012399:Rotation; D025262:Vacuolar Proton-Translocating ATPases / Q000378:metabolism*
chemical_list: D011522:Protons; D000255:Adenosine Triphosphate; D025262:Vacuolar Proton-Translocating ATPases