DOI: 10.1073/pnas.96.14.8173. Secretin PulD: Association with pilot PulS, structure, and ion-conducting channel formation

Secretin PulD: Association with pilot PulS, structure, and ion-conducting channel formation

10.1073/pnas.96.14.8173

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

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.

Bibliography

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, 96(14), 8173â8177.

Authors 7

Nico Nouwen (first)
Neil Ranson (additional)
Helen Saibil (additional)
Bettina Wolpensinger (additional)
Andreas Engel (additional)
Alexandre Ghazi (additional)
Anthony P. Pugsley (additional)

References 26 Referenced 160

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Dates

Type	When
Created	23 years ago (July 26, 2002, 10:38 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 3:37 p.m.)
Indexed	1 year, 3 months ago (May 17, 2024, 11:28 a.m.)
Issued	26 years, 1 month ago (July 6, 1999)
Published	26 years, 1 month ago (July 6, 1999)
Published Online	26 years, 1 month ago (July 6, 1999)
Published Print	26 years, 1 month ago (July 6, 1999)

Funders 0

None

BibTeX

@article{Nouwen_1999, title={Secretin PulD: Association with pilot PulS, structure, and ion-conducting channel formation}, volume={96}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.96.14.8173}, DOI={10.1073/pnas.96.14.8173}, number={14}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Nouwen, Nico and Ranson, Neil and Saibil, Helen and Wolpensinger, Bettina and Engel, Andreas and Ghazi, Alexandre and Pugsley, Anthony P.}, year={1999}, month=jul, pages={8173–8177} }

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