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ADP-ribosylarginine hydrolases

T Takada1, I J Okazaki, J Moss

  • 1Laboratory of Cellular Metabolism, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892.

Molecular and Cellular Biochemistry
|September 1, 1994
PubMed
Summary

ADP-ribosylarginine hydrolase removes ADP-ribose from proteins, reversing ADP-ribosylation. This study identifies and characterizes hydrolase enzymes from various species, revealing conserved and divergent features.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Enzymology

Background:

  • ADP-ribosylation is a crucial reversible post-translational protein modification.
  • NAD:arginine ADP-ribosyltransferase catalyzes ADP-ribose addition to proteins.
  • ADP-ribosylarginine hydrolase reverses this modification by removing ADP-ribose.

Purpose of the Study:

  • To purify and characterize ADP-ribosylarginine hydrolase.
  • To clone and sequence the hydrolase cDNA from rat, mouse, and human.
  • To compare the properties and sequence homology of hydrolases across species.

Main Methods:

  • Protein purification from turkey erythrocytes.
  • Enzyme activity assays with various ADP-ribosylated substrates.
  • cDNA cloning and sequencing from rat, mouse, and human brain.
  • Recombinant protein expression in E. coli.
  • Western blot analysis using anti-hydrolase antibodies.

Main Results:

  • A 39-kDa monomeric ADP-ribosylarginine hydrolase was purified and characterized.
  • The enzyme requires the ADP-ribose moiety for substrate recognition.
  • cDNA sequences revealed high homology between rat, mouse, and human hydrolases.
  • Recombinant rat hydrolase activity is enhanced by Mg2+ and thiol, while human hydrolase is Mg2+-dependent but thiol-independent.
  • Conserved cysteine residues were identified in mammalian hydrolases.

Conclusions:

  • ADP-ribosylarginine hydrolase plays a key role in regulating ADP-ribosylation.
  • Species-specific differences exist in hydrolase activity regulation, particularly regarding thiol dependence.
  • Conserved structural features suggest a conserved function across species, with variations in cofactor requirements.

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