DOI: 10.1126/science.1154994. Reconstitution of Pilus Assembly Reveals a Bacterial Outer Membrane Catalyst

Reconstitution of Pilus Assembly Reveals a Bacterial Outer Membrane Catalyst

10.1126/science.1154994

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

Type 1 pili from uropathogenic Escherichia coli are a prototype of adhesive surface organelles assembled and secreted by the conserved chaperone/usher pathway. We reconstituted type 1 pilus biogenesis from purified pilus proteins. The usher FimD acted as a catalyst to accelerate the ordered assembly of protein subunits independently of cellular energy. Its activity was highly dependent on the adhesin subunit FimH, which triggered the conversion of FimD into a high-efficiency assembly catalyst. Furthermore, a simple kinetic model adequately rationalized usher-catalyzed pilus assembly in vivo. Our results contribute to a mechanistic understanding of protein-catalyzed biogenesis of supramolecular protein complexes at the bacterial outer cell membrane.

Bibliography

Nishiyama, M., Ishikawa, T., Rechsteiner, H., & Glockshuber, R. (2008). Reconstitution of Pilus Assembly Reveals a Bacterial Outer Membrane Catalyst. Science, 320(5874), 376â379.

Authors 4

Mireille Nishiyama (first)
Takashi Ishikawa (additional)
Helene Rechsteiner (additional)
Rudi Glockshuber (additional)

References 27 Referenced 111

10.1038/nrmicro1443
10.1093/emboj/19.12.2803
10.1073/pnas.93.18.9827
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Materials and methods are available as supporting material on Science Online.
10.1016/S0021-9258(18)99895-9
10.1111/j.1365-2958.2007.05657.x
10.1111/j.1574-6968.1996.tb08115.x
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k on = 2 × 10 5 M –1 s –1 k off = 2 s –1 k (A→H) = 4 × 10 –5 s –1 k (A→G) = 8 × 10 –5 s –1 k (A→F) = 5 × 10 –4 s –1 k (A→A) = 16 s –1 . The rate constants k on and k off describe the association with and dissociation from FimD N of FimC-FimA respectively and the rate constants k (A→H) k (A→G) k (A→F) and k (A→A) describe the FimD-mediated DSE reactions between FimA as donor subunit and FimH FimG FimF and FimA as acceptor subunit respectively.
k (A A →H) = 3.0 × 10 –2 ± 0.3 × 10 –2 M –1 s –1 k (A A →G) = 6.5 ± 0.2 M –1 s –1 k (A A →F) = 7.3 ± 0.2 M –1 s –1 k (A A →A) = 2.6 ± 0.1 M –1 s –1 . The rate constants k ( A A → X ) describe the spontaneous DSE reaction between FimA A as donor subunit and the subunit Fim X as acceptor subunit (Fim X is FimH FimG FimF or FimA).
10.1073/pnas.0606795103
We thank K. Hollenstein and L. Thöny-Meyer for constructive discussions and comments on the manuscript and E. Weber-Ban for helpful discussions on kinetic data. We also acknowledge support of the Electron Microscopy Center (EMEZ) of the ETH Zurich. This work was supported by the Swiss National Science Foundation 3100A0-100787 (R.G.) and the ETH Zurich in the framework of the National Center for Competence in Research (NCCR) Structural Biology Program.

Dates

Type	When
Created	17 years, 5 months ago (March 27, 2008, 9:27 p.m.)
Deposited	1 year, 7 months ago (Jan. 10, 2024, 4:24 a.m.)
Indexed	1 day, 7 hours ago (Aug. 29, 2025, 6:13 a.m.)
Issued	17 years, 4 months ago (April 18, 2008)
Published	17 years, 4 months ago (April 18, 2008)
Published Print	17 years, 4 months ago (April 18, 2008)

Funders 0

None

BibTeX

@article{Nishiyama_2008, title={Reconstitution of Pilus Assembly Reveals a Bacterial Outer Membrane Catalyst}, volume={320}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.1154994}, DOI={10.1126/science.1154994}, number={5874}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Nishiyama, Mireille and Ishikawa, Takashi and Rechsteiner, Helene and Glockshuber, Rudi}, year={2008}, month=apr, pages={376–379} }

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