Purification and Characterization of a Complex between Cloacin and Its Immunity Protein Isolated from Enterobacter cloacae (Clo DF13). Dissociation and Reconstitution of the Complex

Graaf, Frits K. de; Klaasen-Boor, P

doi:10.1111/j.1432-1033.1977.tb11296.x

Cited by 59 publications

(44 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, the D/K/K chimera is more active in complex with the ImmK protein (Chi3B) than in its absence (Chi3A). This is in agreement with the empirical observation that the naturally secreted form of the nuclease type colicin-Imm complexes is more toxic than colicin molecules freed in vitro of their Imm proteins (41,42). The C-terminal part of the klebicin D CD is 56.7% identical compared with colicin D (amino acids 422-607), whereas the N-terminal amino acids 314 -421 in colicin D are only weakly similar (27.3% identical residues) (Fig.…”

supporting

confidence: 90%

Dual Roles of the Central Domain of Colicin D tRNase in TonB-mediated Import and in Immunity

Mora

Klepsch

Buckingham

et al. 2008

Journal of Biological Chemistry

View full text Add to dashboard Cite

Colicin D import into Escherichia coli requires an interaction via its TonB box with the energy transducer TonB. Colicin D cytotoxicity is inhibited by specific tonB mutations, but it is restored by suppressor mutations in the TonB box. Here we report that there is a second site of interaction between TonB and colicin D, which is dependent upon a 45-amino acid region, within the uncharacterized central domain of colicin D. In addition, the 8th amino acids of colicin D (a glycine) and colicin B (a valine), adjacent to their TonB boxes, are also required for TonB recognition, suggesting that high affinity complex formation involves multiple interactions between these colicins and TonB. The central domain also contributes to the formation of the immunity complex, as well as being essential for uptake and thus killing. Colicin D is normally secreted in association with the immunity protein, and this complex involves the following two interactions: a major interaction with the C-terminal tRNase domain and a second interaction involving the central domain of colicin D and, most probably, the ␣4 helix of ImmD, which is on the opposite side of ImmD compared with the major interface. In contrast, formation of the immunity complex with the processed cytotoxic domain, the form expected to be found in the cytoplasm after colicin D uptake, requires only the major interaction. Klebicin D has, like colicin D, a ribonuclease activity toward tRNA Arg and a central domain, which can form a complex with ImmD but which does not function in TonB-mediated transport.

show abstract

supporting

confidence: 90%

Dual Roles of the Central Domain of Colicin D tRNase in TonB-mediated Import and in Immunity

Mora

Klepsch

Buckingham

et al. 2008

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…The fact that the producer strains were resistant to acnecin leads us to an expectation that immune mechanisms to endogenous bacteriocin exist in this case as documented for immunity to colicin (13,29) and cloacin (4,20).…”

Section: Methodsmentioning

confidence: 99%

Purification and Properties of a Bacteriocin-Like Substance (Acnecin) of Oral Propionibacterium acnes

Fujimura

Nakamura

1978

Antimicrob Agents Chemother

View full text Add to dashboard Cite

Propionibacterium acnes CN-8, isolated from human dental plaque, was grown in a liquid medium, and its bacteriocin-like substance (acnecin) was extracted from the cells by ultrasonic treatment. Acnecin was purified to a homogeneous state with recovery of 47%. Specific activity increased 72-fold in comparison with the crude extract. The properties of acnecin were as follows. (i) Acnecin may consist of five subunits with a molecular weight of about 12,000. (ii) Its isoelectric point was 5.5. (iii) In amino acid composition, aspartic acid, glutamic acid, glycine, and alanine were predominant, whereas cystine was not present. (iv) Acnecin contained 3.3% carbohydrate but was substantially free from lipid. (v) The activity was lost by heating at 60°C or by protease and lysozyme treatments. Acnecin acted bacteriostatically on the indicator strain without killing it. The action spectrum of acnecin was very narrow; it was effective only against strains of nonacnecin-producing P. acnes and Corynebacterium parvum, a species closely related to P. acnes.

show abstract

“…In addition, IutA binds the bacteriocin cloacin DF13 (16,34) and presumably the bacteriophage B74K (24). Cloacin DF13 is produced by strains of Enterobacter cloacae and secreted as a protein complex comprising a 56-kDa activity protein and a 10-kDa immunity protein (8). Upon interaction with IutA, the immunity protein is released to the medium while cloacin is translocated across the outer membrane (15).…”

mentioning

confidence: 99%

“…Outer membrane protein preparations and immunoblot experiments were conducted by following reported procedures (19,31,32). Cloacin DF13 was purified by column chromatography with CM-Sephadex as previously described (8,18), and its 50% killing activity was determined on susceptible cells (E. coli P9OC cells carrying pJHCV8) grown in M9 minimal medium containing 0.1% bovine serum albumin (9,15). Cloacin sensitivity was evaluated by a plate killing assay in which 0.5 x 108 microorganisms grown overnight in M9 minimal medium were mixed with molten soft agarose containing the iron chelator a, a'-dipyridyl (200 ,M), and the mixture was poured onto L-agar plates.…”

mentioning

confidence: 99%

Role of tol genes in cloacin DF13 susceptibility of Escherichia coli K-12 strains expressing the cloacin DF13-aerobactin receptor IutA

Thomas

Valvano

1993

J Bacteriol

View full text Add to dashboard Cite

IutA is the outer membrane protein receptor for ferric aerobactin and the bacteriocin cloacin DF13. Although the same receptor is shared, ferric aerobactin transport across the outer membrane in Escherichia coli is TonB dependent, whereas cloacin DF13 transport is not. We have recently observed that tolQ is required for cloacin DF13 susceptibility (J. A. Thomas and M. A. Valvano, FEMS Microbiol. Lett. 91:107-112, 1992). In this study, we demonstrate that the genes tolQ, toiR, and tolA, but not tolB, toWC, and ompF, are required for the internalization of cloacin DF13 and they are not involved in the transport of ferric aerobactin.IutA is an iron-regulated outer membrane protein that serves as a receptor for the hydroxamate siderophore ferric aerobactin (6, 16). In addition, IutA binds the bacteriocin cloacin DF13 (16,34) and presumably the bacteriophage B74K (24). Cloacin DF13 is produced by strains of Enterobacter cloacae and secreted as a protein complex comprising a 56-kDa activity protein and a 10-kDa immunity protein (8). Upon interaction with IutA, the immunity protein is released to the medium while cloacin is translocated across the outer membrane (15). Upon entry into the cell cytoplasm, cloacin can block protein synthesis by causing cleavage of the 16S ribosomal RNA (9, 22).Although both ferric aerobactin and cloacin DF13 interact with IutA primarily at the cell surface (34), the translocation of ferric aerobactin across the outer membrane is TonB dependent, whereas that of cloacin DF13 is not (5, 25). TonB, an inner membrane protein with a large periplasmic domain, interacts with various outer membrane receptors and is essential for the transport of ferrisiderophores, certain bacteriophages, the antibiotic albomycin, and group B colicins such as colicins B, G, D, H, and V (see reference 23 for a review). In contrast, group A colicins (colicins A, El, E2, E3, K, L, N, and S4) and filamentous bacteriophages fl, M13, and fd are TonB independent but they require the products of tol genes for translocation across the outer membrane (see reference 35 for a review). The Tol import system consists of various genes such as tolQ, toiR, tolA, tolB, and toiC whose products are located in different cell compartments; i.e., the inner membrane, periplasmic space, and outer membrane (3,26,27,35 Strains and plasmids used are described in Table 1. Purification of plasmid DNA and transformations were carried out as described elsewhere (30). Outer membrane protein preparations and immunoblot experiments were conducted by following reported procedures (19,31,32). Cloacin DF13 was purified by column chromatography with CM-Sephadex as previously described (8, 18), and its 50% killing activity was determined on susceptible cells (E. coli P9OC cells carrying pJHCV8) grown in M9 minimal medium containing 0.1% bovine serum albumin (9, 15). Cloacin sensitivity was evaluated by a plate killing assay in which 0.5 x 108 microorganisms grown overnight in M9 minimal medium were mixed with molten soft agarose containing the iron chela...

show abstract

Purification and Characterization of a Complex between Cloacin and Its Immunity Protein Isolated from Enterobacter cloacae (Clo DF13). Dissociation and Reconstitution of the Complex

Cited by 59 publications

References 25 publications

Dual Roles of the Central Domain of Colicin D tRNase in TonB-mediated Import and in Immunity

Dual Roles of the Central Domain of Colicin D tRNase in TonB-mediated Import and in Immunity

Purification and Properties of a Bacteriocin-Like Substance (Acnecin) of Oral Propionibacterium acnes

Role of tol genes in cloacin DF13 susceptibility of Escherichia coli K-12 strains expressing the cloacin DF13-aerobactin receptor IutA

Contact Info

Product

Resources

About