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• W2148994162 abstract "Abstract Synchronized catalysis in native enzyme : We used a photoactive nanotrigger (NT) to study the initial electron transfer to FAD in native neuronal NOS catalysis. Modeling and fluorescence spectroscopy showed that selective NT binding to NADPH sites is able to override Phe1395 regulation, thus permitting ultrafast injection of electrons into the protein electron pathway. That NT initiation of flavoenzyme catalysis led to the formation of NO is promising for time‐resolved X‐ray and other cellular applications. magnified image Nitric oxide synthases (NOSs) are unique flavohemoproteins with various roles in mammalian physiology. Constitutive NOS catalysis is initiated by fast hydride transfer from NADPH, followed by slower structural rearrangements. We used a photoactive nanotrigger (NT) to study the initial electron transfer to FAD in native neuronal NOS (nNOS) catalysis. Molecular modeling and fluorescence spectroscopy showed that selective NT binding to NADPH sites close to FAD is able to override Phe1395 regulation. Ultrafast injection of electrons into the protein electron pathway by NT photoactivation through the use of a femtosecond laser pulse is thus possible. We show that calmodulin, required for NO synthesis by constitutive NOS, strongly promotes intramolecular electron flow (6.2‐fold stimulation) by a mechanism involving proton transfer to the reduced FAD − site. Site‐directed mutagenesis using the S1176A and S1176T mutants of nNOS supports this hypothesis. The NT synchronized the initiation of flavoenzyme catalysis, leading to the formation of NO, as detected by EPR. This NT is thus promising for time‐resolved X‐ray and other cellular applications." @default.
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• W2148994162 date "2009-02-24" @default.
• W2148994162 modified "2023-10-18" @default.
• W2148994162 title "NO Formation by Neuronal NO-Synthase can be Controlled by Ultrafast Electron Injection from a Nanotrigger" @default.
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• W2148994162 doi "https://doi.org/10.1002/cbic.200800721" @default.
• W2148994162 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/19222033" @default.
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