title: myo-inositol oxygenase from rat kidneys. Substrate-dependent oligomerization.
abstract: myo-Inositol from rat kidneys, an oligomeric protein with apparent molecular mass of about 270 kDa can be dissociated under mild conditions to structured 16.8-kDa monomers. This dissociation can be reversed at high protein concentrations at room temperature. The corresponding apparent dimerization constant K2app = 1.38 x 10(5) M-1, the corresponding rate constant k2 = 350 s-1.M-1, and the apparent constant for the association of dimers, K4app = 2.7 x 10(6) M-1. Reassociation is significantly enhanced in the presence of the substrate and iron(II) (K2app = 9.8 x 10(5) M-1; K4app = 3.75 x 10(6) M-1, k2 = 1750 s-1.M-1, at 20 mM myo-inositol and 0.5 mM FeSO4). Under these conditions almost 100% of the original enzymatic activity was reconstituted. Monomers, with or without bound ligands, lack catalytic activity, whereas the dimer is likely to be the elementary active enzyme-building unit. The effects of myo-inositol on the dimerization lead to the conclusion that this step is both mediated and facilitated by the substrate.
authors.affiliation: Institut für Allgemeine Biochemie, University of Vienna, Austria.
authors.forename: F
authors.initials: F
authors.lastname: Koller
authors.forename: E
authors.initials: E
authors.lastname: Koller
mesh_terms: D000818:Animals; D002850:Chromatography, Gel; D003545:Cysteine / Q000378:metabolism; D005296:Ferrous Compounds / Q000378:metabolism*; D007294:Inositol / Q000378:metabolism*; D050562:Inositol Oxygenase; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D009842:Oligopeptides / Q000096:biosynthesis*; D010105:Oxygenases / Q000378:metabolism*; D011487:Protein Conformation; D011489:Protein Denaturation; D051381:Rats
chemical_list: D005296:Ferrous Compounds; D009842:Oligopeptides; C020748:ferrous sulfate; D007294:Inositol; D010105:Oxygenases; D050562:Inositol Oxygenase; D003545:Cysteine