Screening for nitric oxide-dependent protein-protein interactions.
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.
Matsumoto
Akio
A
Howard Hughes Medical Institute and Department of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
Comatas
Karrie E
KE
Liu
Limin
L
Stamler
Jonathan S
JS
D000818:Animals; D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism*; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004792:Enzyme Precursors / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development; D015723:Gene Library; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D008247:Lysosomes / Q000201:enzymology; D008928:Mitochondria / Q000201:enzymology; D009569:Nitric Oxide / Q000378:metabolism* / Q000494:pharmacology; D020030:Nitric Oxide Donors / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D052248:Nitric Oxide Synthase Type I; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; D010084:Oxidation-Reduction; D011233:Precipitin Tests; D011485:Protein Binding; D015398:Signal Transduction; D013108:Sphingomyelin Phosphodiesterase / Q000378:metabolism*; D014162:Transfection; D014169:Transformation, Bacterial; D014226:Triazenes / Q000494:pharmacology; D020798:Two-Hybrid System Techniques; D001616:beta-Galactosidase / Q000378:metabolism; D019323:omega-N-Methylarginine / Q000494:pharmacology
C105351:1-hydroxy-2-oxo-3,3-bis(2-aminoethyl)-1-triazene; D004791:Enzyme Inhibitors; D004792:Enzyme Precursors; D020030:Nitric Oxide Donors; D014226:Triazenes; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; D006861:Hydrogen Peroxide; C496320:NOS1 protein, human; C496319:NOS2 protein, human; C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052248:Nitric Oxide Synthase Type I; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; D013108:Sphingomyelin Phosphodiesterase; D001616:beta-Galactosidase; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases