. . . "Mart\u00E1sek, Pavel" . "1439-4227" . . "RIV/00216208:11110/09:4029" . "P(KAN200200651), Z(MSM0021620806)" . . "ChemBioChem" . . . . . . "DE - Spolkov\u00E1 republika N\u011Bmecko" . "4" . . "Slama-Schwok, A." . . . "Blanchard-Desce, M." . "Lambry, JC" . . "emission spectroscopy; flavins; kinetics; metalloenzymes; molecular modeling; photocatalysis; nitric-oxide synthase; redox properties; structural basis; 2 flavins; domain; nadph; calmodulin; catalysis; reductase; binding"@en . . "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 S1776T mutants of nNOS supports this hypothesis."@en . "1"^^ . . "NO Formation by Neuronal NO-Synthase can be Controlled by Ultrafast Electron Injection from a Nanotrigger" . . "10" . "[E7FA14346C34]" . . "Brochon, JC" . . "Deprez, E." . "12"^^ . . "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 S1776T mutants of nNOS supports this hypothesis." . "329623" . . . "9"^^ . . . "11110" . "Panda, SP" . "NO Formation by Neuronal NO-Synthase can be Controlled by Ultrafast Electron Injection from a Nanotrigger" . "000264168000014" . "NO Formation by Neuronal NO-Synthase can be Controlled by Ultrafast Electron Injection from a Nanotrigger"@en . "RIV/00216208:11110/09:4029!RIV10-MSM-11110___" . . . . . "van Faassen, EEH" . . "NO Formation by Neuronal NO-Synthase can be Controlled by Ultrafast Electron Injection from a Nanotrigger"@en . . "Beaumont, E." .