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Secretin PulD: Association with pilot PulS, structure, and ion-conducting channel formation
Open accessAbstract
The outer membrane protein PulD (secretin) of Klebsiella oxytoca is required for transport of pullulanase across this membrane. We have purified a multimeric PulD complex from an Escherichia coli strain expressing all the proteins involved in pullulanase secretion. The outer membrane-anchored lipoprotein PulS was found to copurify with PulD. The molar ratio of the two proteins is close to 1:1, and the size of the complex is ≈1 MDa. Scanning transmission electron and cryo-electron microscopy analyses showed that the purified complex is a cylindrical structure having a central cavity of ≈7.6 nm and peripheral radial spokes. Fusion of proteoliposomes containing the purified complex with a planar lipid bilayer resulted in the appearance of small, voltage-activated, ion-conducting channels. We conclude that the central cavity seen in the electron microscope is part of a large gated channel and propose that the observed current fluctuations correspond to voltage-induced, relatively minor displacements of domains in the purified complex rather than to a complete opening of the secretin channel.
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Cited by
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Chen, I., & Dubnau, D. (2004) · Nature Reviews Microbiology · doi:10.1038/nrmicro844 · Cited by 769
The type II secretion system: biogenesis, molecular architecture and mechanism
Korotkov, K. V., Sandkvist, M., & Hol, W. G. J. (2012) · Nature Reviews Microbiology · doi:10.1038/nrmicro2762 · Cited by 549
Recombinant protein secretion in Escherichia coli
Mergulhão, F. J. M., Summers, D. K., & Monteiro, G. A. (2005) · Biotechnology Advances · doi:10.1016/j.biotechadv.2004.11.003 · Cited by 485
Wza the translocon for E. coli capsular polysaccharides defines a new class of membrane protein
Dong, C., Beis, K., Nesper, J., Brunkan-LaMontagne, A. L., Clarke, B. R., Whitfield, C., & Naismith, J. H. (2006) · Nature · doi:10.1038/nature05267 · Cited by 373
Metadata
- Authors
- Nouwen, N., Ranson, N., Saibil, H., Wolpensinger, B., Engel, A., Ghazi, A., & Pugsley, A. P.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 1999-07-06
- Volume
- 96
- Issue
- 14
- Pages
- 8173-8177
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2006668901
- Cited by
- 195
- References
- 0
- FWCI
- 7.60
- Type
- journal article
- Language
- en
- Open access
- green
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- Nouwen, N., Ranson, N., Saibil, H., Wolpensinger, B., Engel, A., Ghazi, A., & Pugsley, A. P. (1999). Secretin PulD: Association with pilot PulS, structure, and ion-conducting channel formation. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.96.14.8173
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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