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Screening for nitric oxide-dependent protein-protein interactions.

Akio Matsumoto·Karrie E Comatas·Limin Liu·Jonathan S Stamler
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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.

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AnimalsApoptosisCaspase 3CaspasesCell LineEnzyme InhibitorsEnzyme PrecursorsEscherichia coliGene LibraryHumansHydrogen PeroxideLysosomesMitochondriaNitric OxideNitric Oxide DonorsNitric Oxide SynthaseNitric Oxide Synthase Type INitric Oxide Synthase Type IINitric Oxide Synthase Type IIIOxidation-ReductionPrecipitin TestsProtein BindingSignal TransductionSphingomyelin PhosphodiesteraseTransfectionTransformation, BacterialTriazenesTwo-Hybrid System Techniquesbeta-Galactosidaseomega-N-MethylarginineJournal Article

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Akio Matsumoto, Karrie E Comatas, Limin Liu, Jonathan S Stamler. 2003-08-01. Screening for nitric oxide-dependent protein-protein interactions.. https://doi.org/10.1126/science.1079319

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