myo-inositol oxygenase from rat kidneys. Substrate-dependent oligomerization

Eur J Biochem. 1990 Oct 24;193(2):421-7. doi: 10.1111/j.1432-1033.1990.tb19355.x.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatography, Gel
  • Cysteine / metabolism
  • Ferrous Compounds / metabolism*
  • Inositol / metabolism*
  • Inositol Oxygenase
  • Kidney / enzymology*
  • Kinetics
  • Oligopeptides / biosynthesis*
  • Oxygenases / metabolism*
  • Protein Conformation
  • Protein Denaturation
  • Rats

Substances

  • Ferrous Compounds
  • Oligopeptides
  • ferrous sulfate
  • Inositol
  • Oxygenases
  • Inositol Oxygenase
  • Cysteine