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Literature summary extracted from

  • Lu, C.; Turley, S.; Marionni, S.T.; Park, Y.J.; Lee, K.K.; Patrick, M.; Shah, R.; Sandkvist, M.; Bush, M.F.; Hol, W.G.
    Hexamers of the type II secretion ATPase GspE from Vibrio cholerae with increased ATPase activity (2013), Structure, 21, 1707-1717.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
7.4.2.8 cardiolipin stimulates activity of full-length GspEEpsE when in complex with the cytoplasmic domain of GspLEpsL Vibrio cholerae
7.4.2.8 additional information hexamerization enhances the ATPase activity of recombinant DN1GspEEpsE-linker-Hcp1 variants compared to monomeric DN1GspEEpsE by more than 20fold Vibrio cholerae

Cloned(Commentary)

EC Number Cloned (Comment) Organism
7.4.2.8 recombinant expression of the enzyme GspE as fusion proteins with Pseudomonas aeruginosa Hcp1, i.e. DELTAN1GspEEpsE-KLASGHcp1, DELTAN1GspEEpsE-GSGSGS-Hcp1, DELTAN1GspEEpsE-KLASGAGHcp1, and DELTAN1GspEEpsE-KLASGAGH-Hcp1 showing homogeneous hexamer formation, several variants of DN1GspEEpsE-Hcp1 fusions with different linker sequences are constructed Vibrio cholerae

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
7.4.2.8 recombinant enzyme GspE-Hcp1 fusion protein hexamers, sitting drop vapour diffusion method, mixing of 0.001 ml protein solution containing 20 mM Tris-HCl, pH 8.0, 500 mM NaCl, 5% v/v glycerol, 1 mM TCEP/HCl, 5 mM AMPPNP, and 5 mM MgCl2 with different precipitant solutions, containing for DN1GspEEpsE-GSGSGS-Hcp1 7% PEG 6000 and 0.1 M bicine, pH 9.0, for DN1GspEEpsE-KLASGAGH-Hcp1 16% PEG 300, 0.2 M ammonium sulfate, 0.1 M Bis Tris, pH 6.1, 5 mM ADP, 5 mM MgCl2, 5 mM AlCl3, and 15 mM NaF, for DELTAN1GspEEpsE-KLASGAG-Hcp1 7% PEG 3350, 0.12 M ammonium citrate pH 7.0, 5 mM ADP, and 5 mM MgCl2, and for DELTAN1GspEEpsE-KLASG-Hcp1 12.5% PEG 20000, 0.1 M bicine pH 9.0, and 2% v/v 1,4-dioxane, at 4°C, X-ray diffraction structure determination and analysis at 4.09-7.6 A resolution Vibrio cholerae

Protein Variants

EC Number Protein Variants Comment Organism
7.4.2.8 additional information four hexamers of Vibrio cholerae GspEEpsE are obtained when fused to Hcp1 as an assistant hexamer, shown with native mass spectrometry. The enzymatic activity of the GspEEpsE-Hcp1 fusions is about 20 times higher than that of a GspEEpsE monomer. Crystal structures of GspEEpsE-Hcp1 fusions with different linker lengths reveal regular and elongated hexamers of GspEEpsE with major differences in domain orientation within subunits, and in subunit assembly Vibrio cholerae

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
7.4.2.8 Mg2+ required Vibrio cholerae

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
7.4.2.8 56000
-
 x * 56000 Vibrio cholerae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
7.4.2.8 ATP + H2O Vibrio cholerae
-
 ADP + phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
7.4.2.8 Vibrio cholerae P37093
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
7.4.2.8 ATP + H2O
-
 Vibrio cholerae ADP + phosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
7.4.2.8 ? x * 56000 Vibrio cholerae
7.4.2.8 More recombinant enzyme GspE-Hcp1 fusion proteins, i.e. DELTAN1GspEEpsE-KLASGHcp1, DELTAN1GspEEpsE-GSGSGS-Hcp1, DELTAN1GspEEpsE-KLASGAGHcp1, and DELTAN1GspEEpsE-KLASGAGH-Hcp1, show homogeneous hexamer formation, structure analysis, detailed overview. The hexamerization enhances the ATPase activity of these four DN1GspEEpsE-linker-Hcp1 variants compared to monomeric DN1GspEEpsE by more than 20fold. The metal binding domains are located on the periphery of both DN1GspEEpsE hexamers Vibrio cholerae

Synonyms

EC Number Synonyms Comment Organism
7.4.2.8 critical multidomain T2SS assembly ATPase
-
 Vibrio cholerae
7.4.2.8 GspE
-
 Vibrio cholerae
7.4.2.8 GspEEpsE
-
 Vibrio cholerae
7.4.2.8 type II secretion ATPase
-
 Vibrio cholerae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
7.4.2.8 37
-
 assay at Vibrio cholerae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4.2.8 8.5
-
 assay at Vibrio cholerae

General Information

EC Number General Information Comment Organism
7.4.2.8 evolution the T2SS secretion ATPase GspE belongs to the family of Type II/IV secretion ATPases Vibrio cholerae
7.4.2.8 metabolism the type II secretion system (T2SS), a multiprotein machinery spanning two membranes in Gram-negative bacteria, incudes the critical multidomain GspEEpsE, and is responsible for the secretion of folded proteins from the periplasm across the outer membrane Vibrio cholerae