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- EMDB-22094: CryoEM structure of the holo-SrpI encapsulin complex from Synecho... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-22094 | |||||||||
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Title | CryoEM structure of the holo-SrpI encapsulin complex from Synechococcus elongatus PCC 7942 | |||||||||
![]() | Cryo-EM Structure of the holo-Srpl encapsulin complex from Synechococcus elongatus PCC 7942 | |||||||||
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![]() | encapsulin / nanocompartment / cysteine desulfurase / HK97-fold / VIRUS LIKE PARTICLE | |||||||||
Function / homology | Type 2A encapsulin shell protein SrpI-like / : / Type 2A encapsulin shell protein SrpI-like / encapsulin nanocompartment / Type 2A encapsulin shell protein SrpI![]() | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.2 Å | |||||||||
![]() | LaFrance BJ / Nichols RJ / Phillips NR / Oltrogge LM / Valentin-Alvarado LE / Bischoff AJ / Savage DF / Nogales E | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Discovery and characterization of a novel family of prokaryotic nanocompartments involved in sulfur metabolism. Authors: Robert J Nichols / Benjamin LaFrance / Naiya R Phillips / Devon R Radford / Luke M Oltrogge / Luis E Valentin-Alvarado / Amanda J Bischoff / Eva Nogales / David F Savage / ![]() ![]() Abstract: Prokaryotic nanocompartments, also known as encapsulins, are a recently discovered proteinaceous organelle-like compartment in prokaryotes that compartmentalize cargo enzymes. While initial studies ...Prokaryotic nanocompartments, also known as encapsulins, are a recently discovered proteinaceous organelle-like compartment in prokaryotes that compartmentalize cargo enzymes. While initial studies have begun to elucidate the structure and physiological roles of encapsulins, bioinformatic evidence suggests that a great diversity of encapsulin nanocompartments remains unexplored. Here, we describe a novel encapsulin in the freshwater cyanobacterium PCC 7942. This nanocompartment is upregulated upon sulfate starvation and encapsulates a cysteine desulfurase enzyme via an N-terminal targeting sequence. Using cryo-electron microscopy, we have determined the structure of the nanocompartment complex to 2.2 Å resolution. Lastly, biochemical characterization of the complex demonstrated that the activity of the cysteine desulfurase is enhanced upon encapsulation. Taken together, our discovery, structural analysis, and enzymatic characterization of this prokaryotic nanocompartment provide a foundation for future studies seeking to understand the physiological role of this encapsulin in various bacteria. #1: ![]() Title: Discovery and characterization of a novel family of prokaryotic nanocompartments involved in sulfur metabolism Authors: Nichols RJ / LaFrance BJ / Phillips NR / Oltrogge LM / Valentin-Alvarado LE / Bischoff AJ / Nogales E / Savage DF | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 53.4 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 21 KB 21 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 18 KB | Display | ![]() |
Images | ![]() | 200.3 KB | ||
Masks | ![]() | 512 MB | ![]() | |
Filedesc metadata | ![]() | 6.5 KB | ||
Others | ![]() ![]() | 412.3 MB 412.3 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6x8mMC ![]() 6x8tC C: citing same article ( M: atomic model generated by this map |
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Similar structure data | |
EM raw data | ![]() Data size: 390.3 Data #1: Gain-corrected raw movies for holo-SrpI encapsulin structure (EMD-22094) [micrographs - multiframe]) |
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | Cryo-EM Structure of the holo-Srpl encapsulin complex from Synechococcus elongatus PCC 7942 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.8625 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
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Projections & Slices |
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Density Histograms |
-Half map: Even half-map for holo-Srpl encapsulin complex
File | emd_22094_half_map_1.map | ||||||||||||
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Annotation | Even half-map for holo-Srpl encapsulin complex | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Odd half-map for holo-Srpl encapsulin complex
File | emd_22094_half_map_2.map | ||||||||||||
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Annotation | Odd half-map for holo-Srpl encapsulin complex | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : SrpI encapsulin holo complex
Entire | Name: SrpI encapsulin holo complex |
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Components |
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-Supramolecule #1: SrpI encapsulin holo complex
Supramolecule | Name: SrpI encapsulin holo complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Molecular weight | Theoretical: 2.1 MDa |
-Supramolecule #2: SrpI encapsulin
Supramolecule | Name: SrpI encapsulin / type: complex / ID: 2 / Parent: 1 / Macromolecule list: all |
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Source (natural) | Organism: ![]() |
-Supramolecule #3: CyD cargo protein
Supramolecule | Name: CyD cargo protein / type: complex / ID: 3 / Parent: 1 |
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Source (natural) | Organism: ![]() |
-Macromolecule #1: Protein SrpI
Macromolecule | Name: Protein SrpI / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() Strain: PCC 7942 / FACHB-805 |
Molecular weight | Theoretical: 35.371121 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MTDNAPQLAL RDVAARQLAN ATKTVPQLRT ITPRWLVRLL HWTPVEAGIY RVNQVKDASQ ITVACSERDE SELPETFVDY IDNPREYLL SAVNTVVDVH TRISDLYSNP HDQIREQLRL TIEIMKERQE SELINSREYG LLNNVAPGQL VHTRNGAPTP D DLDELLIR ...String: MTDNAPQLAL RDVAARQLAN ATKTVPQLRT ITPRWLVRLL HWTPVEAGIY RVNQVKDASQ ITVACSERDE SELPETFVDY IDNPREYLL SAVNTVVDVH TRISDLYSNP HDQIREQLRL TIEIMKERQE SELINSREYG LLNNVAPGQL VHTRNGAPTP D DLDELLIR VWKEPAFFLA HPQAIAAFGR ECTRRGVPPA TVSLFGSSFI TWRGVPLIPS DKVPLENGKT KILLLRVGES RQ GVVGLYQ PNLPGEQGMG LSVRFMGINR KALASYLVSL YCSLAVLTDD ALAVLDNVDV TQYHTYRYNS GHHHHHH UniProtKB: Type 2A encapsulin shell protein SrpI |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 2.6 mg/mL |
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Buffer | pH: 10 |
Grid | Model: Quantifoil, UltrAuFoil, R1.2/1.3 / Material: GOLD / Mesh: 300 Details: Open hole grids were treated with 2 drops of chloroform, and were not glow discharged before applying sample. |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 278 K / Instrument: FEI VITROBOT MARK IV Details: blot force 5, 3 sec blot, 100 humidity, 4C, 1 sec drain time. |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 25 eV |
Software | Name: SerialEM |
Details | Residual beam tilt corrected in RELION/3.1. CTF refinement performed in RELION/3.1. |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Number grids imaged: 1 / Number real images: 1027 / Average electron dose: 35.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.6 mm / Nominal defocus max: 2.8000000000000003 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 64000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Software | Name: PHENIX (ver. dev-2883) |
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Details | One subunit was built completely de novo based on sequence/density, and iteratively real space refined with neighboring subunits using PHENIX. |
Refinement | Space: REAL / Protocol: RIGID BODY FIT / Overall B value: 20 / Target criteria: Correlation coefficient |
Output model | ![]() PDB-6x8m: |