Endogenous ADP-ribosylation of a G(alpha i) protein in bovine ciliary body is stimulated by nitric oxide

Biochem Biophys Res Commun. 1997 Jun 27;235(3):482-6. doi: 10.1006/bbrc.1997.6811.

Abstract

Five ciliary body membrane proteins were labeled when incubated with (adenylate-32P)NAD. Nitric oxide donors stimulated the labeling of 64, 40, and 29-30 kDa proteins and inhibited that of 58 and 56 kDa proteins. The greatest influence of nitric oxide was on the 40 kDa protein: a 17-fold stimulation. Western blotting and immunoprecipitation with specific antibodies identified this protein as the alpha-subunit of G(i-1). Studies with inhibitors showed that the protein was mono-ADP-ribosylated. Treatment of (32P)NAD-labeled G(i-1) with Hg and analysis of the released radioactive material showed that the protein was ADP-ribosylated on a cysteine residue.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Diphosphate Ribose / metabolism
  • Animals
  • Blotting, Western
  • Cattle
  • Cell Membrane / metabolism
  • Ciliary Body / drug effects
  • Ciliary Body / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • GTP-Binding Proteins / isolation & purification
  • GTP-Binding Proteins / metabolism*
  • Hydroxylamine
  • Hydroxylamines / pharmacology
  • Kinetics
  • Macromolecular Substances
  • Membrane Proteins / isolation & purification
  • Membrane Proteins / metabolism*
  • Mercury / pharmacology
  • Molecular Weight
  • NAD / metabolism*
  • Nitric Oxide / pharmacology*
  • Nitrogen Oxides
  • Phosphorus Radioisotopes
  • Spermine / analogs & derivatives*
  • Spermine / pharmacology

Substances

  • Hydroxylamines
  • Macromolecular Substances
  • Membrane Proteins
  • Nitrogen Oxides
  • Phosphorus Radioisotopes
  • NAD
  • spermine nitric oxide complex
  • Adenosine Diphosphate Ribose
  • Hydroxylamine
  • Spermine
  • Nitric Oxide
  • GTP-Binding Proteins
  • Mercury