DOI: 10.1091/mbc.10.7.2163. In Vitro Studies with Purified Components Reveal Signal Recognition Particle (SRP) and SecA/SecB as Constituents of Two Independent Protein-targeting Pathways of<i>Escherichia coli</i>

In Vitro Studies with Purified Components Reveal Signal Recognition Particle (SRP) and SecA/SecB as Constituents of Two Independent Protein-targeting Pathways ofEscherichia coli

10.1091/mbc.10.7.2163

Crossref journal-article

American Society for Cell Biology (ASCB)

Molecular Biology of the Cell (1076)

Abstract

The molecular requirements for the translocation of secretory proteins across, and the integration of membrane proteins into, the plasma membrane of Escherichia coli were compared. This was achieved in a novel cell-free system from E. coliwhich, by extensive subfractionation, was simultaneously rendered deficient in SecA/SecB and the signal recognition particle (SRP) components, Ffh (P48), 4.5S RNA, and FtsY. The integration of two membrane proteins into inside-out plasma membrane vesicles of E. coli required all three SRP components and could not be driven by SecA, SecB, and ΔμH+. In contrast, these were the only components required for the translocation of secretory proteins into membrane vesicles, a process in which the SRP components were completely inactive. Our results, while confirming previous in vivo studies, provide the first in vitro evidence for the dependence of the integration of polytopic inner membrane proteins on SRP in E. coli. Furthermore, they suggest that SRP and SecA/SecB have different substrate specificities resulting in two separate targeting mechanisms for membrane and secretory proteins in E. coli. Both targeting pathways intersect at the translocation pore because they are equally affected by a blocked translocation channel.

Bibliography

Koch, H.-G., Hengelage, T., Neumann-Haefelin, C., MacFarlane, J., Hoffschulte, H. K., Schimz, K.-L., Mechler, B., & MÃ¼ller, M. (1999). In Vitro Studies with Purified Components Reveal Signal Recognition Particle (SRP) and SecA/SecB as Constituents of Two Independent Protein-targeting Pathways ofEscherichia coli. Molecular Biology of the Cell, 10(7), 2163â2173.

Authors 8

Hans-Georg Koch (first)
Thomas Hengelage (additional)
Christoph Neumann-Haefelin (additional)
Juan MacFarlane (additional)
Hedda K. Hoffschulte (additional)
Karl-Ludwig Schimz (additional)
Bernd Mechler (additional)
Matthias Müller (additional)

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Dates

Type	When
Created	12 years, 1 month ago (July 2, 2013, 7:34 p.m.)
Deposited	3 years, 6 months ago (Feb. 26, 2022, 9:39 p.m.)
Indexed	1 month, 1 week ago (July 14, 2025, 11:13 p.m.)
Issued	26 years, 1 month ago (July 1, 1999)
Published	26 years, 1 month ago (July 1, 1999)
Published Print	26 years, 1 month ago (July 1, 1999)

Funders 0

None

BibTeX

@article{Koch_1999, title={In Vitro Studies with Purified Components Reveal Signal Recognition Particle (SRP) and SecA/SecB as Constituents of Two Independent Protein-targeting Pathways ofEscherichia coli}, volume={10}, ISSN={1939-4586}, url={http://dx.doi.org/10.1091/mbc.10.7.2163}, DOI={10.1091/mbc.10.7.2163}, number={7}, journal={Molecular Biology of the Cell}, publisher={American Society for Cell Biology (ASCB)}, author={Koch, Hans-Georg and Hengelage, Thomas and Neumann-Haefelin, Christoph and MacFarlane, Juan and Hoffschulte, Hedda K. and Schimz, Karl-Ludwig and Mechler, Bernd and Müller, Matthias}, editor={Walter, Peter}, year={1999}, month=jul, pages={2163–2173} }

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