Cysteine Scanning Mutagenesis and Disulfide Mapping Analysis of Arrangement of GspC and GspD Protomers within the Type 2 Secretion System

Wang, Xiaohui; Pineau, Camille; Gu, Shuang; Guschinskaya, Natalia; Pickersgill, Richard W.; Shevchik, Vladimir E.

doi:10.1074/jbc.m112.346338

Cited by 38 publications

(64 citation statements)

References 45 publications

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“…7). Alternatively, data from the T2SS suggests that the secretin is assembled as a hexamer of dimers (40)(41)(42); in this case, the arrangement may be one PilMNOP complex for every two PilQ monomers (1:1:1:1:2 stoichiometry). Differences in orientation of the N0 domain of PilQ in a dimer versus monomer configuration might explain why we saw differences in PilPQ interactions when fragments versus full periplasmic domains of PilQ were used for pulldowns.…”

Section: Discussionmentioning

confidence: 99%

PilMNOPQ from the Pseudomonas aeruginosa Type IV Pilus System Form a Transenvelope Protein Interaction Network That Interacts with PilA

et al. 2013

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Section: Discussionmentioning

confidence: 99%

PilMNOPQ from the Pseudomonas aeruginosa Type IV Pilus System Form a Transenvelope Protein Interaction Network That Interacts with PilA

et al. 2013

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“…An important consensus from these studies has been the proposal that monomeric GspD secretins assemble under C 12 symmetry to construct the functional GspD dodecamer. However, several recent studies (17,18) suggested that the assembly of secretin dodecamers might proceed via oligomerization of dimers of secretin subunits.…”

mentioning

confidence: 99%

New Insights into the Assembly of Bacterial Secretins

Meeren

Wen

Gelder

et al. 2013

Journal of Biological Chemistry

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Background: Secretins are outer membrane dodecameric translocation channels in bacterial type II secretion systems (T2SS). Results: The basic assembly unit of XcpQ, the T2SS secretin of the human pathogen Pseudomonas aeruginosa, is a dimer. Conclusion: Functional secretin likely results from hexameric assembly of secretin subunit dimers. Significance: This work is a conceptual advancement in understanding the assembly principles and dynamic function of bacterial secretins.

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“…1A). The N domain is located in the periplasm and interacts with inner membrane components of the secretion machinery (6)(7)(8)(9). The atomic resolution structures of part of the N domains of several secretins have been solved by X-ray crystallography (for the T3SS, EscC [10], and for the T2SS, GspD [11] and XcpQ [12]) and nuclear magnetic resonance (NMR) spectroscopy (for T4P, EscC [13]).…”

mentioning

confidence: 99%

Bacterial Secretins Form Constitutively Open Pores Akin to General Porins

Disconzi¹,

Guilvout²,

Chami³

et al. 2013

Journal of Bacteriology

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e Proteins called secretins form large multimeric complexes that are essential for macromolecular transit across the outer membrane of Gram-negative bacteria. Evidence suggests that the channels formed by some secretin complexes are not tightly closed, but their permeability properties have not been well characterized. Here, we used cell-free synthesis coupled with spontaneous insertion into liposomes to investigate the permeability of the secretin PulD. Leakage assays using preloaded liposomes indicated that PulD allows the efflux of small fluorescent molecules with a permeation cutoff similar to that of general porins. Other secretins were also found to form similar pores. To define the polypeptide region involved in determining the pore size, we analyzed a collection of PulD variants and studied the roles of gates 1 and 2, which were previously reported to affect the pore size of filamentous phage f1 secretin pIV, in assembly and pore formation. Liposome leakage and a novel in vivo assay showed that replacement of the conserved proline residue at position 443 in PulD by leucine increased the apparent size of the pore. The in vitro approach described here could be used to study the pore properties of membrane proteins whose production in vivo is toxic.

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Cysteine Scanning Mutagenesis and Disulfide Mapping Analysis of Arrangement of GspC and GspD Protomers within the Type 2 Secretion System

Cited by 38 publications

References 45 publications

PilMNOPQ from the Pseudomonas aeruginosa Type IV Pilus System Form a Transenvelope Protein Interaction Network That Interacts with PilA

PilMNOPQ from the Pseudomonas aeruginosa Type IV Pilus System Form a Transenvelope Protein Interaction Network That Interacts with PilA

New Insights into the Assembly of Bacterial Secretins

Bacterial Secretins Form Constitutively Open Pores Akin to General Porins

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