Abstract
Because nitric oxide (NO) may be a ubiquitous regulator of cellular signaling, we have modified the yeast two-hybrid system to explore the possibility of NO-dependent protein-protein interactions. We screened for binding partners of procaspase-3, a protein implicated in apoptotic signaling pathways, and identified multiple NO-dependent interactions.Two such interactions, with acid sphingomyelinase and NO synthase, were shown to occur in mammalian cells dependent on endogenous NO. Nitrosylation may thus provide a broad-based mechanism for regulating interactions between proteins. If so, systematic proteomic analyses in which redox state and NO bioavailability are carefully controlled will reveal a large array of novel interactions.
MeSH terms
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Animals
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Apoptosis
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Caspase 3
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Caspases / metabolism*
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Cell Line
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Enzyme Inhibitors / pharmacology
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Enzyme Precursors / metabolism*
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Escherichia coli / genetics
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Escherichia coli / growth & development
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Gene Library
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Humans
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Hydrogen Peroxide / metabolism
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Lysosomes / enzymology
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Mitochondria / enzymology
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Nitric Oxide / metabolism*
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Nitric Oxide / pharmacology
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Nitric Oxide Donors / pharmacology
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Nitric Oxide Synthase / antagonists & inhibitors
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Nitric Oxide Synthase / metabolism*
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Nitric Oxide Synthase Type I
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Nitric Oxide Synthase Type II
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Nitric Oxide Synthase Type III
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Oxidation-Reduction
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Precipitin Tests
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Protein Binding*
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Signal Transduction
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Sphingomyelin Phosphodiesterase / metabolism*
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Transfection
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Transformation, Bacterial
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Triazenes / pharmacology
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Two-Hybrid System Techniques
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beta-Galactosidase / metabolism
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omega-N-Methylarginine / pharmacology
Substances
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1-hydroxy-2-oxo-3,3-bis(2-aminoethyl)-1-triazene
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Enzyme Inhibitors
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Enzyme Precursors
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Nitric Oxide Donors
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Triazenes
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omega-N-Methylarginine
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Nitric Oxide
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Hydrogen Peroxide
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NOS1 protein, human
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NOS2 protein, human
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NOS3 protein, human
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Nitric Oxide Synthase
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Nitric Oxide Synthase Type I
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Nitric Oxide Synthase Type II
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Nitric Oxide Synthase Type III
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Sphingomyelin Phosphodiesterase
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beta-Galactosidase
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CASP3 protein, human
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Caspase 3
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Caspases