myo-Inositol oxygenase: Molecular cloning and expression of a unique enzyme that oxidizes myo-inositol and D-chiro-inositol

R. J. Arner, K. S. Prabhu, J. T. Thompson, G. R. Hildenbrandt, A. D. Liken, C. C. Reddy

Research output: Contribution to journalArticle

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Abstract

myo-Inositol oxygenase (MIOX) catalyses the first committed step in the only pathway of myo-inositol catabolism, which occurs predominantly in the kidney. The enzyme is a non-haemiron enzyme that catalyses the ring cleavage of myo-inositol with the incorporation of a single atom of oxygen. A full-length cDNA was isolated from a pig kidney library with an open reading frame of 849 bp and a corresponding protein subunit molecular mass of 32.7 kDa. The cDNA was expressed in a bacterial pET expression system and an active recombinant MIOX was purified from bacterial lysates to electrophoretic homogeneity. The purified enzyme displayed the same catalytic properties as the native enzyme with Km and kcat values of 5.9 mM and 11 min-1 respectively. The pI was estimated to be 4.5. Preincubation with 1 mM Fe2+ and 2 mM cysteine was essential for the enzyme's activity. D-chiro-Inositol, a myo-inositol isomer, is a substrate for the recombinant MIOX with an estimated Km of 33.5 mM. Both myo-inositol and D-chiro-inositol have been implicated in the pathogenesis of diabetes. Thus an understanding of the regulation of MIOX expression clearly represents a potential window on the aetiology of diabetes as well as on the control of various intracellular phosphoinositides and key signalling pathways.

Original languageEnglish (US)
Pages (from-to)313-320
Number of pages8
JournalBiochemical Journal
Volume360
Issue number2
DOIs
StatePublished - Dec 1 2001

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Inositol Oxygenase
Cloning
Molecular Cloning
Inositol
Enzymes
Medical problems
Complementary DNA
Kidney
Protein Subunits
Enzyme activity
Molecular mass
Phosphatidylinositols
Isomers
Open Reading Frames
Cysteine
Swine
Oxygen
Atoms

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Arner, R. J. ; Prabhu, K. S. ; Thompson, J. T. ; Hildenbrandt, G. R. ; Liken, A. D. ; Reddy, C. C. / myo-Inositol oxygenase : Molecular cloning and expression of a unique enzyme that oxidizes myo-inositol and D-chiro-inositol. In: Biochemical Journal. 2001 ; Vol. 360, No. 2. pp. 313-320.
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myo-Inositol oxygenase : Molecular cloning and expression of a unique enzyme that oxidizes myo-inositol and D-chiro-inositol. / Arner, R. J.; Prabhu, K. S.; Thompson, J. T.; Hildenbrandt, G. R.; Liken, A. D.; Reddy, C. C.

In: Biochemical Journal, Vol. 360, No. 2, 01.12.2001, p. 313-320.

Research output: Contribution to journalArticle

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AU - Arner, R. J.

AU - Prabhu, K. S.

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