Open AccessFunctional reconstitution of a proton-translocating system responsive to fusicoccin.
Author(s) -
Patrizia Aducci,
A. Ballio,
JeanPierre Blein,
Maria Rosaria Fullone,
Michel Rossignol,
René Scalla
Publication year - 1988
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.85.21.7849
Subject(s) - fusicoccin , atpase , biochemistry , chemistry , quenching (fluorescence) , liposome , biophysics , biology , fluorescence , enzyme , physics , quantum mechanics
Crude fusicoccin binding proteins and a partially purified plasma membrane H+-transporting ATPase (EC 3.6.1.34), both solubilized from maize tissues, were simultaneously inserted into liposomes by the freeze-thaw method. ATP-driven intravesicular acidification in the proteoliposomes, measured by the fluorescence quenching of the dye 9-amino-6-chloro-2-methoxyacridine, markedly increased upon addition of fusicoccin to the reconstituted system. This effect could not be observed when binding sites and ATPase preparations were separately reconstituted into the proteoliposomes, thus demonstrating that fusicoccin binding to its receptor is a prerequisite for ATPase stimulation.