TolB
From Proteopedia
StructureTolB is a 44-kDa periplasmic protein partially associated with the outer membrane[1]. It has two domains: an N-terminal α/β domain and a C-terminal six-bladed β-propeller (to which Pal and Colicin E9 bind)[2]. The β-propeller has a latching or ‘Velco’ strand which joins the first and last of the six blades, and is positioned in the domain-domain interface. When Pal binds to the C-terminus of TolB, the latching strand moves away from the interface and carries with it a proline residue. The movement of the latching strand opens up a canyon that would normally be buried between the N- and C-terminal domains of TolB. This canyon can now be used as a binding site for the N-terminal of TolB, which forms a helical half-turn and a β-sheet against the canyon. For additional details see Tol. FunctionThe distal N-terminal 12 residues of TolB has two conformational states which are governed by protein-protein interactions with the β -propeller and results in the binding of TolA in the inner membrane[2]. TolB has been shown to be essential for the function of the Tol system in Escherichia coli[2] by generating an allosteric signal based on a conformational switch in the β-propeller region. TolB has also been shown to interact with the porins of Escherichia coli, in particular OmpF, OmpC, PhoE and LamB, but not OmpA or any of their denatured counterparts. It has been proposed that the whole Tol complex plays a role in this association, although "tol" mutants do not prevent this assembly completely therefore the Tol system may be involved kinetically, not directly[3]. The TolB-Pal ComplexThe TolB-Pal complex is involved in maintaining the outer membrane integrity. Upon binding, TolB and Pal undergo a conformational change , the result of which is crucial for further interactions with other proteins[4]. This complex is parasitised by protein antibiotics and disrupted in order to trigger the translocation of the toxin across the outer membrane (see Colicin for further information)[2]. To study the interaction of TolB with Pal, two studies were carried out[1]: TolBBep (tagging TolB for immunoprecipitation), which allows the associated proteins to remain in contact with TolB, and in vivo cross-linking experiments with formaldehyde. Immunoprecipitation gave the result that Pal co-precipitates with TolBBep, while the cross-linking showed that in the present of Pal, the two products migrated close to each other, but in the absence of Pal, neither band was present, demonstrating an interaction between the two. These two experiments showed that TolB directly interacts with Pal, and that this interaction is responsible for maintaining the association of TolB with the membrane.
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3D structures of TolB
Updated on 25-April-2022
1crz, 1c5k – EcTolB – Escherichia coli
2ivz – EcTolB + colicin-E9 T-domain
4jml - EcTolB (mutant) + colicin-E9 T-domain (mutant)
7nst - EcTolB (mutant) + colicin-E9 + outer membrane protein F – Cryo EM
7nsu - EcTolB (mutant) + colicin-E9 + outer membrane protein F + BtuB – Cryo EM
3iax - EcTolB + colicin-A translocation domain
2hqs, 2w8b - EcTolB + peptidoglycan-associated lipoprotein
4pwz – YpTolB – Yersinia pestis
4r40 - YpTolB + peptidoglycan-associated lipoprotein
6pnv – TolB – Salmonella enterica
7mx5 – TolB – Acinetobacteria baumannii
References
- ↑ 1.0 1.1 Bouveret E, Derouiche R, Rigal A, Lloubes R, Lazdunski C, Benedetti H. Peptidoglycan-associated lipoprotein-TolB interaction. A possible key to explaining the formation of contact sites between the inner and outer membranes of Escherichia coli. J Biol Chem. 1995 May 12;270(19):11071-7. PMID:7744736
- ↑ 2.0 2.1 2.2 2.3 Bonsor DA, Hecht O, Vankemmelbeke M, Sharma A, Krachler AM, Housden NG, Lilly KJ, James R, Moore GR, Kleanthous C. Allosteric beta-propeller signalling in TolB and its manipulation by translocating colicins. EMBO J. 2009 Sep 16;28(18):2846-57. Epub 2009 Aug 20. PMID:19696740 doi:10.1038/emboj.2009.224
- ↑ Rigal A, Bouveret E, Lloubes R, Lazdunski C, Benedetti H. The TolB protein interacts with the porins of Escherichia coli. J Bacteriol. 1997 Dec;179(23):7274-9. PMID:9393690
- ↑ Godlewska R, Wisniewska K, Pietras Z, Jagusztyn-Krynicka EK. Peptidoglycan-associated lipoprotein (Pal) of Gram-negative bacteria: function, structure, role in pathogenesis and potential application in immunoprophylaxis. FEMS Microbiol Lett. 2009 Sep;298(1):1-11. Epub 2009 May 21. PMID:19519769 doi:10.1111/j.1574-6968.2009.01659.x
