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- EMDB-2729: Conformational Snapshots of Inducible Nitric Oxide Synthase (iNOS) -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-2729 | |||||||||
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Title | Conformational Snapshots of Inducible Nitric Oxide Synthase (iNOS) | |||||||||
![]() | Single-particle reconstruction of iNOS: Group I, Conformation xii | |||||||||
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![]() | heterogeneity / random conical tilt / nitric oxide synthase / calmodulin / heme / electron transfer / flavin | |||||||||
Function / homology | ![]() Nitric oxide stimulates guanylate cyclase / ROS and RNS production in phagocytes / G protein-coupled receptor signaling pathway coupled to cGMP nucleotide second messenger / Peroxisomal protein import / cAMP-dependent protein kinase regulator activity / positive regulation of killing of cells of another organism / prostaglandin secretion / tetrahydrobiopterin binding / arginine binding / cortical cytoskeleton ...Nitric oxide stimulates guanylate cyclase / ROS and RNS production in phagocytes / G protein-coupled receptor signaling pathway coupled to cGMP nucleotide second messenger / Peroxisomal protein import / cAMP-dependent protein kinase regulator activity / positive regulation of killing of cells of another organism / prostaglandin secretion / tetrahydrobiopterin binding / arginine binding / cortical cytoskeleton / superoxide metabolic process / cGMP-mediated signaling / nitric-oxide synthase binding / peptidyl-cysteine S-nitrosylation / regulation of cytokine production involved in inflammatory response / cellular response to cytokine stimulus / regulation of insulin secretion / cellular response to organic cyclic compound / nitric-oxide synthase (NADPH) / blood vessel remodeling / nitric oxide mediated signal transduction / response to tumor necrosis factor / nitric-oxide synthase activity / arginine catabolic process / nitric oxide biosynthetic process / negative regulation of blood pressure / response to hormone / positive regulation of interleukin-8 production / response to bacterium / Hsp90 protein binding / negative regulation of protein catabolic process / cellular response to type II interferon / regulation of blood pressure / beta-catenin binding / circadian rhythm / positive regulation of interleukin-6 production / cellular response to xenobiotic stimulus / FMN binding / peroxisome / NADP binding / flavin adenine dinucleotide binding / actin binding / regulation of cell population proliferation / cellular response to lipopolysaccharide / response to lipopolysaccharide / response to hypoxia / calmodulin binding / intracellular signal transduction / defense response to bacterium / inflammatory response / cadherin binding / iron ion binding / positive regulation of apoptotic process / negative regulation of gene expression / heme binding / protein kinase binding / perinuclear region of cytoplasm / protein homodimerization activity / extracellular space / identical protein binding / nucleus / metal ion binding / plasma membrane / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / negative staining / Resolution: 73.0 Å | |||||||||
![]() | Campbell MG / Smith BC / Potter CS / Carragher B / Marletta MA | |||||||||
![]() | ![]() Title: Molecular architecture of mammalian nitric oxide synthases. Authors: Melody G Campbell / Brian C Smith / Clinton S Potter / Bridget Carragher / Michael A Marletta / ![]() Abstract: NOSs are homodimeric multidomain enzymes responsible for producing NO. In mammals, NO acts as an intercellular messenger in a variety of signaling reactions, as well as a cytotoxin in the innate ...NOSs are homodimeric multidomain enzymes responsible for producing NO. In mammals, NO acts as an intercellular messenger in a variety of signaling reactions, as well as a cytotoxin in the innate immune response. Mammals possess three NOS isoforms--inducible, endothelial, and neuronal NOS--that are composed of an N-terminal oxidase domain and a C-terminal reductase domain. Calmodulin (CaM) activates NO synthesis by binding to the helical region connecting these two domains. Although crystal structures of isolated domains have been reported, no structure is available for full-length NOS. We used high-throughput single-particle EM to obtain the structures and higher-order domain organization of all three NOS holoenzymes. The structures of inducible, endothelial, and neuronal NOS with and without CaM bound are similar, consisting of a dimerized oxidase domain flanked by two separated reductase domains. NOS isoforms adopt many conformations enabled by three flexible linkers. These conformations represent snapshots of the continuous electron transfer pathway from the reductase domain to the oxidase domain, which reveal that only a single reductase domain participates in electron transfer at a time, and that CaM activates NOS by constraining rotational motions and by directly binding to the oxidase domain. Direct visualization of these large conformational changes induced during electron transfer provides significant insight into the molecular underpinnings governing NO formation. | |||||||||
History |
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Structure visualization
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 52.8 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 10 KB 10 KB | Display Display | ![]() |
Images | ![]() | 33.7 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 2718C ![]() 2719C ![]() 2720C ![]() 2721C ![]() 2722C ![]() 2723C ![]() 2724C ![]() 2725C ![]() 2726C ![]() 2727C ![]() 2728C ![]() 2730C ![]() 2731C ![]() 2732C ![]() 2733C ![]() 2734C ![]() 2735C ![]() 2736C ![]() 2737C ![]() 2738C ![]() 2739C ![]() 2740C ![]() 2741C ![]() 2742C ![]() 2743C ![]() 2744C ![]() 2745C ![]() 2746C ![]() 2747C ![]() 2748C ![]() 2749C C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Single-particle reconstruction of iNOS: Group I, Conformation xii | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.36 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : Murine Inducible Nitric Oxide Synthase
Entire | Name: Murine Inducible Nitric Oxide Synthase |
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Components |
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-Supramolecule #1000: Murine Inducible Nitric Oxide Synthase
Supramolecule | Name: Murine Inducible Nitric Oxide Synthase / type: sample / ID: 1000 / Details: Sample is highly flexible / Oligomeric state: Homodimer / Number unique components: 1 |
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Molecular weight | Theoretical: 260 KDa |
-Macromolecule #1: Inducible Nitric Oxide Synthase
Macromolecule | Name: Inducible Nitric Oxide Synthase / type: protein_or_peptide / ID: 1 / Name.synonym: iNOS / Number of copies: 1 / Oligomeric state: Homodimer / Recombinant expression: Yes |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 260 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | UniProtKB: Nitric oxide synthase, inducible GO: nitric oxide biosynthetic process, FMN binding, NADP binding, calmodulin binding, flavin adenine dinucleotide binding, heme binding, iron ion binding, nitric-oxide synthase activity InterPro: Ser/Thr protein kinase, TGFB receptor |
-Experimental details
-Structure determination
Method | negative staining |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.5 / Details: 50 mM TEA pH 7.5, 150 mM NaCl, and 5 mM DTT |
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Staining | Type: NEGATIVE Details: 3 microliters of sample were applied to grid. The specimen was stained twice with 2% uranyl formate, then allowed to air-dry. |
Grid | Details: Glow discharged C-flat grid with 2-micron-diameter holes overlaid by thin 1.5 nm continuous carbon |
Vitrification | Cryogen name: NONE / Instrument: OTHER |
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Electron microscopy
Microscope | FEI TECNAI F20 |
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Date | Mar 6, 2013 |
Image recording | Category: CCD / Film or detector model: TVIPS TEMCAM-F416 (4k x 4k) / Number real images: 2226 / Average electron dose: 37 e/Å2 Details: Each area is imaged twice: once at 55 degrees and again with no tilt. |
Tilt angle min | 0 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Calibrated magnification: 114705 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 62000 |
Sample stage | Specimen holder model: SIDE ENTRY, EUCENTRIC / Tilt angle max: 55 |
Experimental equipment | ![]() Model: Tecnai F20 / Image courtesy: FEI Company |
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Image processing
Details | See publication. |
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CTF correction | Details: Each Image |
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 73.0 Å / Resolution method: OTHER / Software - Name: Appion, Spider / Details: See publication. / Number images used: 179 |
Final two d classification | Number classes: 1 |