Effect of C-terminal truncation on the molecular chaperone function and dimerization of Escherichia coli trigger factor
- PMID: 16380200
- DOI: 10.1016/j.biochi.2005.11.006
Effect of C-terminal truncation on the molecular chaperone function and dimerization of Escherichia coli trigger factor
Abstract
To examine the role of the C-terminal domain in the chaperone function of trigger factor (TF), a number of truncation mutants were constructed, namely: TF419, TF389, TF380, TF360, TF344, and TF251, in which the C-terminal 13, 43, 52, 72, 88 residues or the entire C-domain were deleted, respectively. Co-expression of mutant chicken adenylate kinase (AK) with TF and the C-terminal truncation mutants was achieved using a plasmid pBVAT that allows expression of TF and AK from a single plasmid. The results show that truncation of the C-terminus of TF has only minor effect on its ability to assist AK refolding in vivo. Further, ribosome-binding experiments indicate that C-terminal truncation mutants can still bind to the ribosome and the presence of the C-terminus may in fact lower the affinity of TF for the ribosome in vivo. This indicates that the C-domain of trigger factor may not be essential for the ribosome-associated molecular chaperone function of TF. However, the purified TF C-terminal truncation mutants had a dramatically reduced ability to assist rabbit muscle GAPDH refolding in vitro and a reduced tendency to dimerize. This shows that the structural integrity of the C-terminus contributes to both the chaperone function of TF and the stability of the dimeric form.
Similar articles
-
PPIase domain of trigger factor acts as auxiliary chaperone site to assist the folding of protein substrates bound to the crevice of trigger factor.Int J Biochem Cell Biol. 2010 Jun;42(6):890-901. doi: 10.1016/j.biocel.2010.01.019. Epub 2010 Jan 21. Int J Biochem Cell Biol. 2010. PMID: 20096367
-
Functional dissection of Escherichia coli trigger factor: unraveling the function of individual domains.J Bacteriol. 2004 Jun;186(12):3777-84. doi: 10.1128/JB.186.12.3777-3784.2004. J Bacteriol. 2004. PMID: 15175291 Free PMC article.
-
Identification of a potential hydrophobic peptide binding site in the C-terminal arm of trigger factor.Protein Sci. 2007 Jun;16(6):1165-75. doi: 10.1110/ps.062623707. Protein Sci. 2007. PMID: 17525465 Free PMC article.
-
A cradle for new proteins: trigger factor at the ribosome.Curr Opin Struct Biol. 2005 Apr;15(2):204-12. doi: 10.1016/j.sbi.2005.03.005. Curr Opin Struct Biol. 2005. PMID: 15837180 Review.
-
Ribosome-tethered molecular chaperones: the first line of defense against protein misfolding?Curr Opin Microbiol. 2003 Apr;6(2):157-62. doi: 10.1016/s1369-5274(03)00030-4. Curr Opin Microbiol. 2003. PMID: 12732306 Review.
Cited by
-
Protein folding in vitro and in the cell: From a solitary journey to a team effort.Biophys Chem. 2022 Aug;287:106821. doi: 10.1016/j.bpc.2022.106821. Epub 2022 Apr 29. Biophys Chem. 2022. PMID: 35667131 Free PMC article. Review.
-
The dynamic dimer structure of the chaperone Trigger Factor.Nat Commun. 2017 Dec 8;8(1):1992. doi: 10.1038/s41467-017-02196-7. Nat Commun. 2017. PMID: 29222465 Free PMC article.
-
Trigger Factor in Association with the ClpP1P2 Heterocomplex of Leptospira Promotes Protease/Peptidase Activity.ACS Omega. 2021 Jan 7;6(2):1400-1409. doi: 10.1021/acsomega.0c05057. eCollection 2021 Jan 19. ACS Omega. 2021. PMID: 33490799 Free PMC article.
-
Trigger factor from the psychrophilic bacterium Psychrobacter frigidicola is a monomeric chaperone.J Bacteriol. 2009 Feb;191(4):1162-8. doi: 10.1128/JB.01137-08. Epub 2008 Dec 5. J Bacteriol. 2009. PMID: 19060145 Free PMC article.
-
Single-molecule dynamics of the molecular chaperone trigger factor in living cells.Mol Microbiol. 2016 Dec;102(6):992-1003. doi: 10.1111/mmi.13529. Epub 2016 Sep 30. Mol Microbiol. 2016. PMID: 27626893 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Molecular Biology Databases
Research Materials
Miscellaneous
