title: Oligomeric structure of H(+)-translocating inorganic pyrophosphatase of plant vacuoles.
abstract: The topography and oligomeric structure of the vacuolar membrane-bound inorganic pyrophosphatase (73,000 daltons) of mung bean were studied. When the vacuolar membranes were treated with thiocyanate or sodium carbonate which are known to remove the peripheral membrane proteins, the enzyme could not be detected in the solubilized fraction by the specific antibody. The apparent molecular size of the enzyme was estimated to be about 480 kDa by polyacrylamide gel electrophoresis in the presence of Triton X-100. Crosslinking treatment of the pyrophosphatase with dimethyl suberimidate produced a complex corresponding to the dimer. The rate of PPi hydrolysis showed a sigmoidal relationship to substrate concentration with a Hill coefficient of 2.5. These results suggest that the vacuolar pyrophosphatase is an integral membrane protein and functions as an oligomer, probably a dimer.
authors.affiliation: Institute of Low Temperature Science, Hokkaido University, Sapporo, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Maeshima
mesh_terms: D002254:Carbonates / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D007887:Fabaceae / Q000201:enzymology*; D006868:Hydrolysis; D043564:Inorganic Pyrophosphatase; D008565:Membrane Proteins / Q000302:isolation & purification; D008970:Molecular Weight; D009842:Oligopeptides / Q000302:isolation & purification; D010946:Plants, Medicinal; D011487:Protein Conformation; D011755:Pyrophosphatases / Q000302:isolation & purification; D013861:Thiocyanates / Q000494:pharmacology; D014617:Vacuoles / Q000201:enzymology*
chemical_list: D002254:Carbonates; D008565:Membrane Proteins; D009842:Oligopeptides; D013861:Thiocyanates; C005686:sodium carbonate; D011755:Pyrophosphatases; D043564:Inorganic Pyrophosphatase