The X-ray Structure of the Type II Secretion System Complex Formed by the N-terminal Domain of EpsE and the Cytoplasmic Domain of EpsL of Vibrio cholerae

Abendroth, Jan; Murphy, Patricia; Sandkvist, Maria; Bagdasarian, Michael; Hól, Wim G. J.

doi:10.1016/j.jmb.2005.02.061

Cited by 93 publications

(155 citation statements)

References 36 publications

Supporting

Mentioning

145

Contrasting

Order By: Relevance

“…results), as well as pull-down assay (Chen et al, 2005). Co-crystallization of the N-terminal region of EpsE and the cytoplasmic domain of EpsL (Abendroth et al, 2005) further supports the proposition that the N-terminal region of GspE interacts directly with the cytoplasmic domain of GspL. Shiue et al (2006) recently demonstrated that the X. campestris pv.…”

Section: Introductionsupporting

confidence: 55%

Mutation of a key residue in the type II secretion system ATPase uncouples ATP hydrolysis from protein translocation

Shiue

Chien²,

Chan³

et al. 2007

Molecular Microbiology

View full text Add to dashboard Cite

SummaryMembrane-associated ATPase constitutes an essential element common to all secretion machineries in Gram-negative bacteria. How ATP hydrolysis by these ATPases is coupled to secretion process remains unclear. Here we identified R286 as a key residue in the type II secretion system (T2SS) ATPase XpsE of Xanthomonas campestris that plays a pivotal role in coupling ATP hydrolysis to protein translocation. Mutation of R286 to alanine made XpsE hydrolyse ATP at a rate five times that of the wild-type XpsE. Yet the mutant XpsE(R286A) is non-functional in protein secretion via T2SS. Detailed analyses indicated that the mutant XpsE(R286A) lost the ability co-ordinating the N-and C-domain of XpsE. Without significantly influencing XpsE binding affinity with ATP or its oligomerization, R286A mutation however, caused XpsE lose the ability to associate with the cytoplasmic membrane via XpsL N. As a consequence, ATP hydrolysis by XpsE was uncoupled from protein secretion. Because R286 is highly conserved among members of the secretion NTPase superfamily, we speculate that its equivalent in other homologues may also play a critical energy coupling role for T2SS, type IV pilus assembly and type IV secretion system.

show abstract

Section: Introductionsupporting

confidence: 55%

Mutation of a key residue in the type II secretion system ATPase uncouples ATP hydrolysis from protein translocation

Shiue

Chien²,

Chan³

et al. 2007

Molecular Microbiology

View full text Add to dashboard Cite

show abstract

“…Fleckenstein et al (2000) also noted similarity to EpsE, the V. cholerae member of type II/type IV secretion NTPases that provide energy for bacterial protein secretion (Camberg & Sandkvist, 2005). EpsE is known to have both a cytoplasmic (Abendroth et al, 2005) and innermembrane association (Camberg & Sandkvist, 2005), dependent upon the expression of other Eps proteins.…”

Section: Resultsmentioning

confidence: 96%

Biochemical characterization of the enterotoxigenic Escherichia coli LeoA protein

Brown

Hardwidge

2007

Microbiology

View full text Add to dashboard Cite

Enterotoxigenic Escherichia coli (ETEC) causes enterotoxin-induced diarrhoea and significant mortality. The molecular mechanisms underlying how the heat-labile enterotoxin (LT) is secreted during infection are poorly understood. ETEC produce outer-membrane vesicles (OMVs) containing LT that are endocytosed into host cells. Although OMV production and protein content may be a regulated component of ETEC pathogenesis, how LT loading into OMVs is regulated is unknown. The LeoA protein plays a role in secreting LT from the bacterial periplasm. To begin to understand the function of LeoA and its role in ETEC H10407 pathogenesis, a site-directed mutant lacking the putative GTP-binding domain was constructed. The ability of wild-type and mutant LeoA to hydrolyse GTP in vitro was quantified. This domain was found to be responsible for GTP binding; it is important to LeoA's function in LT secretion, and may play a modest role in the formation and protein content of OMVs. Deletion of leoA reduced the abundance of OmpX in outer-membrane protein preparations and of LT in OMVs. Immunoprecipitation experiments revealed that LeoA interacts directly with OmpA, but that the GTP-binding domain is nonessential for this interaction. Deletion of leoA rendered ETEC H10407 non-motile, through apparent periplasmic accumulation of FliC.

show abstract

“…Abendroth et al determined the crystal structure of a complex between the N-terminal domain of the T2SS ATPase EpsE and the cytoplasmic domain of EpsL (40), which is similar in structure to PilM (11). In addition, BfpC, a key protein in the biogenesis of type IVb bundle forming pili, shares structural similarity with EpsL and PilM (41) and binds to its cognate assembly ATPase, BfpD (40,42). Interestingly, both the cytoplasmic and periplasmic domains of EpsL form dimers (24,43), raising the question of how this is consistent with binding to the hexameric EpsE ATPase (44).…”

Section: Discussionmentioning

confidence: 99%

Structure and assembly of an inner membrane platform for initiation of type IV pilus biogenesis

Karuppiah

Collins

Thistlethwaite

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Significance Type IV pili are long, thin fibers, formed mainly of polymers of a single pilin protein, which are displayed on the surfaces of many bacteria, including several human pathogens. Here, we report three-dimensional reconstructions of the PilMNO inner membrane complex, alone and in complex with pilin protein, through a combination of X-ray crystallography and electron microscopy. PilMNO forms a dimeric T-shaped structure, binding two copies of the pilin protein at its extremities. The results provide a structural model for the way in which pilin is harvested from the inner membrane and made available to other components of the type IV pilus biogenesis machinery.

show abstract

The X-ray Structure of the Type II Secretion System Complex Formed by the N-terminal Domain of EpsE and the Cytoplasmic Domain of EpsL of Vibrio cholerae

Cited by 93 publications

References 36 publications

Mutation of a key residue in the type II secretion system ATPase uncouples ATP hydrolysis from protein translocation

Mutation of a key residue in the type II secretion system ATPase uncouples ATP hydrolysis from protein translocation

Biochemical characterization of the enterotoxigenic Escherichia coli LeoA protein

Structure and assembly of an inner membrane platform for initiation of type IV pilus biogenesis

Contact Info

Product

Resources

About