N

NADH peroxidase cysteine-sulfenic acid absorbance spectroscopy monitoring, 50-51

microenvironment, 46, 54 preservation in crystal studies, 49-50 reactivity, 45, 48 redox scheme, 45-46 verification in proteins, 47-48 X-ray crystallography crystallization, 49-50 modeling oxidized form, 51-52 reduced form, 52-53 NADH-reduced structure, 52 NADPH oxidase antisense oligodeoxyribonucleotide inhibition activity assay, 430 advantages, 434 controls, 425

delivery to cells, 426-427 effectiveness of inhibition, 429-432 monocyte isolation, 427-428 phosphorothioate modifications of nucleotides, 425-426 purification of oligodeoxyribonucleotides,

426 rationale, 423 sequence design, 423-424 superoxide assay, 428 Western blot of protein expression, 429-431 assay of nonphagocytic enzyme homogenate preparation cultured cells, 220-221 tissue, 221 membrane preparation, 221-222 NADPH consumption, 222-223 pyridine nucleotide dose response, 222 superoxide production aconitase, 232-233 chemiluminescence, 226-227 cytochrome c reduction, 225-226 2',7'-dichlorofluorescein diacetate,

227-229 dihydroethidium, 227-230 electron spin resonance, 231-232 nitroblue tetrazolium, 230-231 phagocytic oxidase considerations, 224-225 catalytic reaction, 220 expression regulation by signaling proteins phospholipase A2, 432-433 protein kinase C, 432-433 inhibition of nonphagocytic enzyme antisense inhibition, 223-224 inhibitors, 223 knockout mice, 224 leukocyte components, 421-422 low-density lipoprotein oxidation by monocytes, 422

phagocytic versus nonphagocytic enzyme comparison, 220-221 respiratory burst, 421 NAP-110, nitric oxide synthase interactions,

238-240 Neelaredoxin complementation analysis in Escherichia coli, 151-152

cytochrome c reduction assay, 150-151 expression analysis in parasite, 153-154 gene, 148 orthologs, 148

oxidation with superoxide, 151 purification from recombinant Escherichia coli, 148-149 reduction using rubredoxin, 151,154 superoxide reductase activity, 141, 149-151 Neonatal stroke, hypoxia-ischemia model copper/zinc-superoxide dismutase transgenic mouse overexpression adult versus neonatal findings, 390-391 enzyme assays in brain catalase, 394

glutathione peroxidase, 394 hydrogen peroxide, 394-395 histology brain sample preparation, 391-392 cresyl violet staining, 392 Perl's iron stain, 392-393 hydrogen peroxide accumulation in neurotoxicity, 396-397 hypoxia-ischemia model modifications, 390

neuron culture studies cell culture, 395-396 overview, 395 hypoxia, 390 overview, 389 surgery, 389-390 NF-kB, see Nuclear factor-)cB Nicotinamide adenine dinucleotide chromatin remodeling role, 282-283, 291 measurement in intact yeast, 299-300 metabolism, 282 oxidase, see NADPH oxidase peroxidase, see NADH peroxidase Nitric oxide olfaction role, 219

protein modification, see S-nitrosylation Nitric oxide synthase catalytic mechanism, 114-115, 117,119 huntingtin transgenic mouse activity assay,

371-373 protein-protein interactions coimmunoprecipitation, 237-238 examples, 233-234

glutathione S-transferase fusion protein pull-down assay, 237 isoform specificity, 233-234 NAP-110 cotransfection studies, 238-240 yeast two-hybrid system bait plasmid, 236 complementary DNA library preparation, 235-236 principles, 235 screening, 236 tetrahydrobiopterin low-temperature optical absorption spectral detection advantages and limitations, 120-121 cry osol vent, 116 equipment, 116

interpretation of results, 117, 119 P450 enzyme comparison, 119 rationale, 115

sample preparation, 116-117 spectra acquisition, 117 stopped-flow data comparison, 120 Nitroblue tetrazolium, superoxide assay for

NADPH oxidase detection, 230-231 Nitrosylation, see S-nitrosylation Northern blot chemokine analysis, 466 cyclic nucleotide-gated channel expression, 217

glutathione peroxidase distribution in transgenic mice, 348-349 manganese superoxide dismutase transgenic mouse RNA, 402 NOS, see Nitric oxide synthase Nuclear factor-/cB

anti-nuclear factor-«: B drug receptor purification with affinity beads, 85-86 electrophoretic mobility shift assay, 470-471 Western blot of I-kB, 471, 473-474

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