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
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Adenosine Diphosphate Ribose / metabolism
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Animals
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Blotting, Western
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Cattle
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Cell Membrane / metabolism
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Ciliary Body / drug effects
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Ciliary Body / metabolism*
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Electrophoresis, Polyacrylamide Gel
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GTP-Binding Proteins / isolation & purification
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GTP-Binding Proteins / metabolism*
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Hydroxylamine
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Hydroxylamines / pharmacology
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Kinetics
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Macromolecular Substances
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Membrane Proteins / isolation & purification
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Membrane Proteins / metabolism*
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Mercury / pharmacology
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Molecular Weight
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NAD / metabolism*
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Nitric Oxide / pharmacology*
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Nitrogen Oxides
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Phosphorus Radioisotopes
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Spermine / analogs & derivatives*
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Spermine / pharmacology
Substances
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Hydroxylamines
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Macromolecular Substances
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Membrane Proteins
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Nitrogen Oxides
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Phosphorus Radioisotopes
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NAD
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spermine nitric oxide complex
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Adenosine Diphosphate Ribose
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Hydroxylamine
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Spermine
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Nitric Oxide
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GTP-Binding Proteins
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Mercury