title: Hexamers of the type II secretion ATPase GspE from Vibrio cholerae with increased ATPase activity.
abstract: The type II secretion system (T2SS), a multiprotein machinery spanning two membranes in Gram-negative bacteria, is responsible for the secretion of folded proteins from the periplasm across the outer membrane. The critical multidomain T2SS assembly ATPase GspE(EpsE) had not been structurally characterized as a hexamer. Here, four hexamers of Vibrio cholerae GspE(EpsE) are obtained when fused to Hcp1 as an assistant hexamer, as shown with native mass spectrometry. The enzymatic activity of the GspE(EpsE)-Hcp1 fusions is ∼20 times higher than that of a GspE(EpsE) monomer, indicating that increasing the local concentration of GspE(EpsE) by the fusion strategy was successful. Crystal structures of GspE(EpsE)-Hcp1 fusions with different linker lengths reveal regular and elongated hexamers of GspE(EpsE) with major differences in domain orientation within subunits, and in subunit assembly. SAXS studies on GspE(EpsE)-Hcp1 fusions suggest that even further variability in GspE(EpsE) hexamer architecture is likely.
authors: Department of Biochemistry, University of Washington, Seattle, WA 98195, USA.
authors: Connie
authors: C
authors: Lu
authors: Stewart
authors: S
authors: Turley
authors: Samuel T
authors: ST
authors: Marionni
authors: Young-Jun
authors: YJ
authors: Park
authors: Kelly K
authors: KK
authors: Lee
authors: Marcella
authors: M
authors: Patrick
authors: Ripal
authors: R
authors: Shah
authors: Maria
authors: M
authors: Sandkvist
authors: Matthew F
authors: MF
authors: Bush
authors: Wim G J
authors: WG
authors: Hol
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry*; D000255:Adenosine Triphosphate / Q000737:chemistry; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D058947:Bacterial Secretion Systems; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D006868:Hydrolysis; D007700:Kinetics; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D011550:Pseudomonas aeruginosa / Q000201:enzymology; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D053838:Scattering, Small Angle; D014734:Vibrio cholerae / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems; D011993:Recombinant Fusion Proteins; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases
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