Catalysis on dinuclear Mn(II) centers

Hydrolytic and redox activities of rat liver arginase

Thomas M. Sossong, Sergei V. Khangulov, R. Christopher Cavalli, Dianne Robert Soprano, G Charles Dismukes, David E. Ash

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Rat liver arginase contains a dinuclear Mn2(II,II) center in each subunit having EPR properties similar to those observed in Mn-catalases. The principal physiologic role of arginase is catalyzing the hydrolytic cleavage of L-arginine to produce L-ornithine and urea. Here we demonstrate that arginase catalyzes the disproportionation of hydrogen peroxide by a redox mechanism analogous to Mn-catalases, but at rates that are 10-5 to 10-6 of k(cat) for the Mn-catalases, and also exhibits peroxidase activity. The dinuclear Mn2(II,II) center is essential for maximal catalase activity, since both the H101N and H126N mutant arginases containing only one Mn(II)/subunit have catalase activities that are <3% of that for the wild- type enzyme. Like the Mn-catalases, the catalase activity of arginase is not inhibited by millimolar concentrations of CN-, the most potent inhibitor of heme catalases, or by EDTA, a chelator of free metal ions. The catalase activity of arginase is not significantly inhibited by C1- or F-, in contrast to Mn-catalases, while potent inhibitors of the hydrolytic activity are also effective inhibitors of the catalase activity. These results suggest that lower affinity of hydrogen peroxide to the active site of arginase contributes to the lower catalase activity. EPR spectroscopy reveals that potent inhibitors of the hydrolytic reaction, including N(ω)-hydroxy-L- arginine, L-lysine, and L-valine, decouple the electronic interaction between the Mn2+ ions, most probably by removing a μ-bridging ligand or by increasing the intermanganese separation. The capacity for arginase to deliver a hydroxide ion to hydrolyze the L-arginine substrate is suggested to arise from a 'dinuclear effect', wherein the two metal ions contribute more or less equivalently in deprotonation of metal-bound water molecule. Structure-reactivity analyses of these reactions will provide insights into the factors that control redox versus hydrolytic function in dimanganese clusters.

Original languageEnglish
Pages (from-to)433-443
Number of pages11
JournalJournal of Biological Inorganic Chemistry
Volume2
Issue number4
DOIs
Publication statusPublished - Aug 1997

Fingerprint

Arginase
Catalysis
Liver
Catalase
Oxidation-Reduction
Rats
Arginine
Metals
Ions
Hydrogen Peroxide
Metal ions
Paramagnetic resonance
Deprotonation
Ornithine
Valine
Chelating Agents
Heme
Edetic Acid
Peroxidase
Lysine

Keywords

  • Arginase
  • Catalase
  • Manganoenzymes
  • Peroxidase

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Catalysis on dinuclear Mn(II) centers : Hydrolytic and redox activities of rat liver arginase. / Sossong, Thomas M.; Khangulov, Sergei V.; Cavalli, R. Christopher; Soprano, Dianne Robert; Dismukes, G Charles; Ash, David E.

In: Journal of Biological Inorganic Chemistry, Vol. 2, No. 4, 08.1997, p. 433-443.

Research output: Contribution to journalArticle

Sossong, Thomas M. ; Khangulov, Sergei V. ; Cavalli, R. Christopher ; Soprano, Dianne Robert ; Dismukes, G Charles ; Ash, David E. / Catalysis on dinuclear Mn(II) centers : Hydrolytic and redox activities of rat liver arginase. In: Journal of Biological Inorganic Chemistry. 1997 ; Vol. 2, No. 4. pp. 433-443.
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abstract = "Rat liver arginase contains a dinuclear Mn2(II,II) center in each subunit having EPR properties similar to those observed in Mn-catalases. The principal physiologic role of arginase is catalyzing the hydrolytic cleavage of L-arginine to produce L-ornithine and urea. Here we demonstrate that arginase catalyzes the disproportionation of hydrogen peroxide by a redox mechanism analogous to Mn-catalases, but at rates that are 10-5 to 10-6 of k(cat) for the Mn-catalases, and also exhibits peroxidase activity. The dinuclear Mn2(II,II) center is essential for maximal catalase activity, since both the H101N and H126N mutant arginases containing only one Mn(II)/subunit have catalase activities that are <3{\%} of that for the wild- type enzyme. Like the Mn-catalases, the catalase activity of arginase is not inhibited by millimolar concentrations of CN-, the most potent inhibitor of heme catalases, or by EDTA, a chelator of free metal ions. The catalase activity of arginase is not significantly inhibited by C1- or F-, in contrast to Mn-catalases, while potent inhibitors of the hydrolytic activity are also effective inhibitors of the catalase activity. These results suggest that lower affinity of hydrogen peroxide to the active site of arginase contributes to the lower catalase activity. EPR spectroscopy reveals that potent inhibitors of the hydrolytic reaction, including N(ω)-hydroxy-L- arginine, L-lysine, and L-valine, decouple the electronic interaction between the Mn2+ ions, most probably by removing a μ-bridging ligand or by increasing the intermanganese separation. The capacity for arginase to deliver a hydroxide ion to hydrolyze the L-arginine substrate is suggested to arise from a 'dinuclear effect', wherein the two metal ions contribute more or less equivalently in deprotonation of metal-bound water molecule. Structure-reactivity analyses of these reactions will provide insights into the factors that control redox versus hydrolytic function in dimanganese clusters.",
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AU - Soprano, Dianne Robert

AU - Dismukes, G Charles

AU - Ash, David E.

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N2 - Rat liver arginase contains a dinuclear Mn2(II,II) center in each subunit having EPR properties similar to those observed in Mn-catalases. The principal physiologic role of arginase is catalyzing the hydrolytic cleavage of L-arginine to produce L-ornithine and urea. Here we demonstrate that arginase catalyzes the disproportionation of hydrogen peroxide by a redox mechanism analogous to Mn-catalases, but at rates that are 10-5 to 10-6 of k(cat) for the Mn-catalases, and also exhibits peroxidase activity. The dinuclear Mn2(II,II) center is essential for maximal catalase activity, since both the H101N and H126N mutant arginases containing only one Mn(II)/subunit have catalase activities that are <3% of that for the wild- type enzyme. Like the Mn-catalases, the catalase activity of arginase is not inhibited by millimolar concentrations of CN-, the most potent inhibitor of heme catalases, or by EDTA, a chelator of free metal ions. The catalase activity of arginase is not significantly inhibited by C1- or F-, in contrast to Mn-catalases, while potent inhibitors of the hydrolytic activity are also effective inhibitors of the catalase activity. These results suggest that lower affinity of hydrogen peroxide to the active site of arginase contributes to the lower catalase activity. EPR spectroscopy reveals that potent inhibitors of the hydrolytic reaction, including N(ω)-hydroxy-L- arginine, L-lysine, and L-valine, decouple the electronic interaction between the Mn2+ ions, most probably by removing a μ-bridging ligand or by increasing the intermanganese separation. The capacity for arginase to deliver a hydroxide ion to hydrolyze the L-arginine substrate is suggested to arise from a 'dinuclear effect', wherein the two metal ions contribute more or less equivalently in deprotonation of metal-bound water molecule. Structure-reactivity analyses of these reactions will provide insights into the factors that control redox versus hydrolytic function in dimanganese clusters.

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