+Open data
-Basic information
Entry | Database: PDB / ID: 7w6x | |||||||||||||||
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Title | Crystal structure of E. coli RseP in complex with batimastat | |||||||||||||||
Components | Regulator of sigma-E protease RseP | |||||||||||||||
Keywords | HYDROLASE / Intramembrane protease | |||||||||||||||
Function / homology | Function and homology information anti-sigma factor antagonist activity / Hydrolases; Acting on peptide bonds (peptidases); Metalloendopeptidases / cellular response to cell envelope stress / metalloendopeptidase activity / positive regulation of DNA-templated transcription / proteolysis / metal ion binding / plasma membrane Similarity search - Function | |||||||||||||||
Biological species | Escherichia coli (E. coli) | |||||||||||||||
Method | X-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 3.2 Å | |||||||||||||||
Authors | Takanuki, K. / Imaizumi, Y. / Nogi, T. | |||||||||||||||
Funding support | Japan, 4items
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Citation | Journal: Sci Adv / Year: 2022 Title: Mechanistic insights into intramembrane proteolysis by site-2 protease homolog RseP. Authors: Yuki Imaizumi / Kazunori Takanuki / Takuya Miyake / Mizuki Takemoto / Kunio Hirata / Mika Hirose / Rika Oi / Tatsuya Kobayashi / Kenichi Miyoshi / Rie Aruga / Tatsuhiko Yokoyama / Shizuka ...Authors: Yuki Imaizumi / Kazunori Takanuki / Takuya Miyake / Mizuki Takemoto / Kunio Hirata / Mika Hirose / Rika Oi / Tatsuya Kobayashi / Kenichi Miyoshi / Rie Aruga / Tatsuhiko Yokoyama / Shizuka Katagiri / Hiroaki Matsuura / Kenji Iwasaki / Takayuki Kato / Mika K Kaneko / Yukinari Kato / Michiko Tajiri / Satoko Akashi / Osamu Nureki / Yohei Hizukuri / Yoshinori Akiyama / Terukazu Nogi / Abstract: Site-2 proteases are a conserved family of intramembrane proteases that cleave transmembrane substrates to regulate signal transduction and maintain proteostasis. Here, we elucidated crystal ...Site-2 proteases are a conserved family of intramembrane proteases that cleave transmembrane substrates to regulate signal transduction and maintain proteostasis. Here, we elucidated crystal structures of inhibitor-bound forms of bacterial site-2 proteases including RseP. Structure-based chemical modification and cross-linking experiments indicated that the RseP domains surrounding the active center undergo conformational changes to expose the substrate-binding site, suggesting that RseP has a gating mechanism to regulate substrate entry. Furthermore, mutational analysis suggests that a conserved electrostatic linkage between the transmembrane and peripheral membrane-associated domains mediates the conformational changes. In vivo cleavage assays also support that the substrate transmembrane helix is unwound by strand addition to the intramembrane β sheet of RseP and is clamped by a conserved asparagine residue at the active center for efficient cleavage. This mechanism underlying the substrate binding, i.e., unwinding and clamping, appears common across distinct families of intramembrane proteases that cleave transmembrane segments. | |||||||||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 7w6x.cif.gz | 117.6 KB | Display | PDBx/mmCIF format |
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PDB format | pdb7w6x.ent.gz | 75.2 KB | Display | PDB format |
PDBx/mmJSON format | 7w6x.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7w6x_validation.pdf.gz | 832.7 KB | Display | wwPDB validaton report |
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Full document | 7w6x_full_validation.pdf.gz | 835.9 KB | Display | |
Data in XML | 7w6x_validation.xml.gz | 17.8 KB | Display | |
Data in CIF | 7w6x_validation.cif.gz | 23.1 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/w6/7w6x ftp://data.pdbj.org/pub/pdb/validation_reports/w6/7w6x | HTTPS FTP |
-Related structure data
Related structure data | 7w6yC 7w6zC 7w70C 7w71C C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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1 |
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Unit cell |
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-Components
#1: Protein | Mass: 50103.047 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Escherichia coli (E. coli) / Strain: K12 / Gene: rseP, ecfE, yaeL, b0176, JW0171 / Production host: Escherichia coli (E. coli) References: UniProt: P0AEH1, Hydrolases; Acting on peptide bonds (peptidases); Metalloendopeptidases | ||||
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#2: Chemical | #3: Chemical | ChemComp-BAT / | Has ligand of interest | Y | |
-Experimental details
-Experiment
Experiment | Method: X-RAY DIFFRACTION / Number of used crystals: 1 |
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-Sample preparation
Crystal | Density Matthews: 3.17 Å3/Da / Density % sol: 61.2 % |
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Crystal grow | Temperature: 293 K / Method: lipidic cubic phase / pH: 7 Details: 30% (vol./vol.) polyethylene glycol 500 dimethyl ether, 100 mM NaCl, 100 mM MgCl2, 100 mM HEPES-Na (pH 7.0) |
-Data collection
Diffraction | Mean temperature: 100 K / Serial crystal experiment: N |
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Diffraction source | Source: SYNCHROTRON / Site: SPring-8 / Beamline: BL32XU / Wavelength: 1 Å |
Detector | Type: DECTRIS EIGER X 9M / Detector: PIXEL / Date: Dec 14, 2018 |
Radiation | Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray |
Radiation wavelength | Wavelength: 1 Å / Relative weight: 1 |
Reflection | Resolution: 3.2→43.77 Å / Num. obs: 10129 / % possible obs: 99.7 % / Redundancy: 8.4 % / Biso Wilson estimate: 103.16 Å2 / CC1/2: 0.995 / Rpim(I) all: 0.098 / Net I/σ(I): 7.5 |
Reflection shell | Resolution: 3.2→3.31 Å / Mean I/σ(I) obs: 1.1 / Num. unique obs: 1028 / CC1/2: 0.417 / Rpim(I) all: 1.347 |
-Processing
Software |
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Refinement | Method to determine structure: MOLECULAR REPLACEMENT / Resolution: 3.2→43.76 Å / SU ML: 0.521 / Cross valid method: FREE R-VALUE / σ(F): 1.93 / Phase error: 33.9463 Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2
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Solvent computation | Shrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL | ||||||||||||||||||||||||||||
Displacement parameters | Biso mean: 94.52 Å2 | ||||||||||||||||||||||||||||
Refinement step | Cycle: LAST / Resolution: 3.2→43.76 Å
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Refine LS restraints |
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LS refinement shell |
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