Crystal structure of Colicin Ia

S-964 Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0448, USA.
Nature (Impact Factor: 41.46). 02/1997; 385(6615):461-4. DOI: 10.1038/385461a0
Source: PubMed


The ion-channel forming colicins A, B, E1, Ia, Ib and N all kill bacterial cells selectively by co-opting bacterial active-transport pathways and forming voltage-gated ion conducting channels across the plasma membrane of the target bacterium. The crystal structure of colicin Ia reveals a molecule 210 A long with three distinct functional domains arranged along a backbone of two extraordinarily long alpha-helices. A central domain at the bend of the hairpin-like structure mediates specific recognition and binding to an outer-membrane receptor. A second domain mediates translocation across the outer membrane via the TonB transport pathway; the TonB-box recognition element of colicin Ia is on one side of three 80 A-long helices arranged as a helical sheet. A third domain is made up of 10 alpha-helices which form a voltage-activated and voltage-gated ion conducting channel across the plasma membrane of the target cell. The two 160 A-long alpha-helices that link the receptor-binding domain to the other domains enable the colicin Ia molecule to span the periplasmic space and contact both the outer and plasma membranes simultaneously during function.

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    • "Using whole colicin N or purified R or TR domains bound Fig. 1. (A) Colicin Ia crystal structure (Wiener et al., 1997), with T domain in blue, R domain in green and channel-forming domain in red. The same colour scheme is used for colicin N (B), but only a single residue of the 92-residue T domain is seen in this structure (Vetter et al., 1998). "
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    ABSTRACT: The mechanisms by which colicins, protein toxins produced by Escherichia coli, kill other E. coli, have become much better understood in recent years. Most colicins initially bind to an outer membrane protein receptor, and then search for a separate nearby outer membrane protein translocator that serves as a pathway into target cells. Many colicins use the outer membrane porin, OmpF, as that translocator, while using a different primary receptor. Colicin N is unique among known colicins in that only OmpF had been identified as being required for uptake of the colicin and it was presumed to somehow serve as both receptor and translocator. Genetic screens also identified a number of genes required for lipopolysaccharide (LPS) synthesis as uniquely required for killing by colicin N, but not by other colicins. In this issue of Molecular Microbiology, Johnson et al. show that the receptor-binding domain of colicin N binds to LPS, and does not require OmpF for that binding. LPS of a minimal length is required for binding, explaining the requirement for specific elements of the LPS biosynthetic pathway. For colicin N, the receptor-binding domain does not recognize a protein, but rather the most abundant component of the outer membrane itself, LPS.
    Molecular Microbiology 03/2014; 92(3). DOI:10.1111/mmi.12569 · 4.42 Impact Factor
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    • "It is within the IUTD that periplasmic Ton or Tol-binding epitopes are typically located. A. ColIa bound to its primary receptor Cir R through its R-domain (Wiener et al., 1997; Buchanan et al., 2007). The structured TonB boxes of Cir proteins are shown as molecular surfaces (orange). "
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    ABSTRACT: The ability of Escherichia coli to kill other E. coli using protein antibiotics known as colicins has been known for many years, but the mechanisms involved poorly understood. Recent progress has been rapid, however, particularly concerning events on either side of the outer membrane (OM). Structures of colicins bound to OM receptors have been determined and we have detailed mechanistic information on how colicins subvert the periplasmic complexes of TolQRAB/Pal or TonB/ExbB/ExbD to trigger cell entry. In this issue of Molecular Microbiology, Jakes and Finkelstein answer a long-standing problem concerning the uptake mechanism of the pore-forming colicin ColIa: How does the TonB box of the colicin cross the OM following high-affinity binding of ColIa to its primary receptor, the siderophore transporter Cir? Through a series of chimeric protein constructions tested for their activity against a range of mutants and in cell death protection assays, the authors come up with the surprising observation that following binding of ColIa to Cir it recruits another Cir protein as its OM translocator. Not only does this settle various conundrums in the literature, but the translocation mechanism that stems from their study will likely be applicable to many TonB-dependent colicins.
    Molecular Microbiology 02/2010; 75(3):529-33. DOI:10.1111/j.1365-2958.2009.06967.x · 4.42 Impact Factor
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    • "Schematic diagram of domain structure of colicins Ia and E3, the chimeric construct IaE3R, the R domain deletion of colicin Ia, IaΔR, and the C-terminal channel-forming domain of colicin Ia, CT-M. Crystal structures of colicin Ia (Wiener et al., 1997) and colicin E3 (Soelaiman et al., 2001) are also shown, with the domains colored as in the schematic diagrams. The R domain of colicin Ia that was replaced or deleted comprises residues 251-407; the R domain of colicin E3 that replaced the Ia R domain included residues 312 to 447 of colicin E3. "
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    ABSTRACT: Colicin Ia, a channel-forming bactericidal protein, uses the outer membrane protein, Cir, as its primary receptor. To kill Escherichia coli, it must cross this membrane. The crystal structure of Ia receptor-binding domain bound to Cir, a 22-stranded plugged beta-barrel protein, suggests that the plug does not move. Therefore, another pathway is needed for the colicin to cross the outer membrane, but no 'second receptor' has ever been identified for TonB-dependent colicins, such as Ia. We show that if the receptor-binding domain of colicin Ia is replaced by that of colicin E3, this chimera effectively kills cells, provided they have the E3 receptor (BtuB), Cir, and TonB. This is consistent with wild-type Ia using one Cir as its primary receptor (BtuB in the chimera) and a second Cir as the translocation pathway for its N-terminal translocation (T) domain and its channel-forming C-terminal domain. Deletion of colicin Ia's receptor-binding domain results in a protein that kills E. coli, albeit less effectively, provided they have Cir and TonB. We show that purified T domain competes with Ia and protects E. coli from being killed by it. Thus, in addition to binding to colicin Ia's receptor-binding domain, Cir also binds weakly to its translocation domain.
    Molecular Microbiology 11/2009; 75(3):567-78. DOI:10.1111/j.1365-2958.2009.06966.x · 4.42 Impact Factor
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