In vitro analysis of the process of translocation of OmpA across the Escherichia coli cytoplasmic membrane. A translocation intermediate accumulates transiently in the absence of the proton motive force - PubMed (original) (raw)
. 1989 Nov 5;264(31):18582-8.
Affiliations
- PMID: 2553715
Free article
In vitro analysis of the process of translocation of OmpA across the Escherichia coli cytoplasmic membrane. A translocation intermediate accumulates transiently in the absence of the proton motive force
K Tani et al. J Biol Chem. 1989.
Free article
Abstract
The proton motive force (delta mu H+) plays an important role, although it is not absolutely essential, in the in vitro translocation of secretory proteins, such as OmpA, across the cytoplasmic membrane of Escherichia coli (Yamada, H., Tokuda, H., and Mizushima, S. (1989) J. Biol. Chem. 264, 1723-1728). The transient accumulation in membrane vesicles of a possible translocation intermediate of OmpA was observed in the absence of delta mu H+. The intermediate was detected on a polyacrylamide gel as a proteinase K-resistant band corresponding to a molecular weight of 26,000. The intermediate did not possess the signal peptide. The appearance of this band was inhibited in the absence of ATP or the presence of adenosine 5'-(beta,gamma-imino)triphosphate (AMP-PNP) and enhanced upon the addition of SecA. Upon the addition of NADH that energizes the membrane, the intermediate was converted to the translocated form of OmpA, even in the presence of AMP-PNP. These results suggest different requirements of ATP and delta mu H+ for the early and late stages of the translocation reaction. The SecA requirement for the early stage of the translocation has also been suggested. In addition to this band, two other bands were observed at higher positions on the gel, when the translocation reaction was performed in the absence of delta mu H+. Although these two bands also represented the mature form of OmpA, which was partly protected from the proteinase K treatment by the membrane vesicles, the accumulation was not transient. These bands did not appear when the translocation reaction was performed in the presence of dithiothreitol. Together with other evidence, the above observations suggest that OmpA, which has an intramolecular disulfide bridge, cannot undergo the translocation unless delta mu H+ is imposed.
Similar articles
- Bacterial protein translocation: kinetic and thermodynamic role of ATP and the protonmotive force.
Driessen AJ. Driessen AJ. Trends Biochem Sci. 1992 Jun;17(6):219-23. doi: 10.1016/0968-0004(92)90381-i. Trends Biochem Sci. 1992. PMID: 1502724 Review. - Export and sorting of the Escherichia coli outer membrane protein OmpA.
Freudl R, Klose M, Henning U. Freudl R, et al. J Bioenerg Biomembr. 1990 Jun;22(3):441-9. doi: 10.1007/BF00763176. J Bioenerg Biomembr. 1990. PMID: 2202726 Review.
Cited by
- A unifying mechanism for protein transport through the core bacterial Sec machinery.
Allen WJ, Collinson I. Allen WJ, et al. Open Biol. 2023 Aug;13(8):230166. doi: 10.1098/rsob.230166. Epub 2023 Aug 30. Open Biol. 2023. PMID: 37643640 Free PMC article. Review. - How Quality Control Systems AID Sec-Dependent Protein Translocation.
Jiang C, Wynne M, Huber D. Jiang C, et al. Front Mol Biosci. 2021 Apr 13;8:669376. doi: 10.3389/fmolb.2021.669376. eCollection 2021. Front Mol Biosci. 2021. PMID: 33928127 Free PMC article. Review. - A High-Resolution Luminescent Assay for Rapid and Continuous Monitoring of Protein Translocation across Biological Membranes.
Pereira GC, Allen WJ, Watkins DW, Buddrus L, Noone D, Liu X, Richardson AP, Chacinska A, Collinson I. Pereira GC, et al. J Mol Biol. 2019 Apr 5;431(8):1689-1699. doi: 10.1016/j.jmb.2019.03.007. Epub 2019 Mar 13. J Mol Biol. 2019. PMID: 30878481 Free PMC article. - Coassembly of SecYEG and SecA Fully Restores the Properties of the Native Translocon.
Bariya P, Randall LL. Bariya P, et al. J Bacteriol. 2018 Dec 7;201(1):e00493-18. doi: 10.1128/JB.00493-18. Print 2019 Jan 1. J Bacteriol. 2018. PMID: 30275279 Free PMC article. - The way is the goal: how SecA transports proteins across the cytoplasmic membrane in bacteria.
Cranford-Smith T, Huber D. Cranford-Smith T, et al. FEMS Microbiol Lett. 2018 Jun 1;365(11):fny093. doi: 10.1093/femsle/fny093. FEMS Microbiol Lett. 2018. PMID: 29790985 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Molecular Biology Databases
Research Materials
Miscellaneous