Absence of periplasmic DsbA oxidoreductase facilitates export of cysteine-containing passenger proteins to the Escherichia coli cell surface via the Igaβ autotransporter pathway

Jose, Joachim; Krämer, Joachim; Klauser, Thomas; Pohlner, Johannes; Meyer, Thomas F.

doi:10.1016/0378-1119(96)00343-5

Cited by 93 publications

(89 citation statements)

References 10 publications

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“…It was proposed that the chaperone activity of DegP was involved (410). It is worth mentioning here that the formation of disulfide bonds in the passenger domain in the presence of the periplasmic disulfide bond-forming enzyme DsbA decreases the efficiency of secretion of the passenger domain (46,232,499,510). This suggests that the proprotein is accessible to periplasmic enzymes and that at least partial folding of the autotransporter may arise in the periplasm.…”

Section: Autotransporter Secretion Pathway (Type Va)mentioning

confidence: 83%

Type V Protein Secretion Pathway: the Autotransporter Story

Henderson

Navarro-Garcı́a

Desvaux

et al. 2004

Microbiol Mol Biol Rev

712

795

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Gram-negative bacteria possess an outer membrane layer which constrains uptake and secretion of solutes and polypeptides. To overcome this barrier, bacteria have developed several systems for protein secretion. The type V secretion pathway encompasses the autotransporter proteins, the two-partner secretion system, and the recently described type Vc or AT-2 family of proteins. Since its discovery in the late 1980s, this family of secreted proteins has expanded continuously, due largely to the advent of the genomic age, to become the largest group of secreted proteins in gram-negative bacteria. Several of these proteins play essential roles in the pathogenesis of bacterial infections and have been characterized in detail, demonstrating a diverse array of function including the ability to condense host cell actin and to modulate apoptosis. However, most of the autotransporter proteins remain to be characterized. In light of new discoveries and controversies in this research field, this review considers the autotransporter secretion process in the context of the more general field of bacterial protein translocation and exoprotein function

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Section: Autotransporter Secretion Pathway (Type Va)mentioning

confidence: 83%

Type V Protein Secretion Pathway: the Autotransporter Story

Henderson

Navarro-Garcı́a

Desvaux

et al. 2004

Microbiol Mol Biol Rev

712

795

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“…In common with all other autotransporter proteins thus far described, AspA from meningococcal strains Z2491, MC58, and Z4181 contains few (five) cysteine residues. It is postulated that a high cysteine content may interfere with translocation through the outer membrane due to the formation of disulfide bonds (18). Of the five cysteine residues present in AspA, one is located at the predicted N terminus of the mature peptide and the remaining four are present as two pairs.…”

Section: Discussionmentioning

confidence: 99%

Autotransported Serine Protease A of Neisseria meningitidis : an Immunogenic, Surface-Exposed Outer Membrane, and Secreted Protein

2002

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Several autotransporter proteins have previously been identified in Neisseria meningitidis. Using molecular features common to most members of the autotransporter family of proteins, we have identified an additional novel ca. 112-kDa autotransporter protein in the meningococcal genomic sequence data. This protein, designated autotransported serine protease A (AspA), has significant N-terminal homology to the secreted serine proteases (subtilases) from several organisms and contains a serine protease catalytic triad. The amino acid sequence of AspA is well-conserved in serogroup A, B, and C meningococci. In Neisseria gonorrhoeae, the AspA homologue appears to be a pseudogene. The gene encoding AspA was cloned and expressed from meningococcal strain MC58 (B15:P1.16b). Anti-AspA antibodies were detected in patients' convalescent-phase sera, suggesting that AspA is expressed in vivo during infection and is immunogenic and cross-reactive. Rabbit polyclonal monospecific anti-AspA serum was used to probe whole-cell proteins from a panel of wild-type meningococcal strains and two AspA mutant strains. Expression of the ca. 112-kDa precursor polypeptide was detected in 12 of 20 wild-type meningococcal strains examined, suggesting that AspA expression is phase variable. Immunogold electron microscopy and cellular fractionation studies showed that the AspA precursor is transported to the outer membrane and remains surface exposed. Western blot experiments confirmed that smaller, ca. 68-or 70-kDa components of AspA (AspA68 and AspA70, respectively) are then secreted into the meningococcal culture supernatant. Site-directed mutagenesis of S426 abolished secretion of both rAspA68 and rAspA70 in Escherichia coli, confirming that AspA is an autocleaved autotransporter protein. In conclusion, we characterized a novel, surface-exposed and secreted, immunogenic, meningococcal autotransporter protein.

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“…These passenger proteins included CTB (19,20,21,24), pseudoazurin of Alcaligenes faecalis (36), MalE and PhoA (38), and various defined epitopes (24; C. T. Lattemann, unpublished data). Display of functional protein domains has not been demonstrated for this system until recently (39).…”

Section: Discussionmentioning

confidence: 99%

“…A limitation, however, is the incompatibility for the translocation of passenger domains containing extensive tertiary structures such as disulfide bonds (20). This restriction could be overcome by inactivating the dsbA gene product of E. coli, leading to the export of wild-type CTB fused to the autotransporter domain of the IgA1 protease of N. gonorrhoeae in the dsbA strain JK321 (19).…”

Section: Discussionmentioning

confidence: 99%

“…The outer membrane protease OmpT is known to degrade heterologous passenger proteins displayed by autotransporters on the cell surface (21,24), and the presence of the periplasmic oxidoreductase DsbA, required for the efficient formation of disulfide bonds in the periplasm (1), has been shown to hamper the translocation of passenger domains containing stable tertiary structures that contain disulfide bonds (19). Interestingly, full-length FP77 was expressed in all E. coli strains examined (Fig.…”

Section: Fig 2 (A)mentioning

confidence: 99%

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Autodisplay: Functional Display of Active β-Lactamase on the Surface of Escherichia coli by the AIDA-I Autotransporter

Lattemann

Maurer

Gerland³

et al. 2000

J Bacteriol

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Members of the protein family of immunoglobulin A1 protease-like autotransporters comprise multidomain precursors consisting of a C-terminal autotransporter domain that promotes the translocation of N-terminally attached passenger domains across the cell envelopes of gram-negative bacteria. Several autotransporter domains have recently been shown to efficiently promote the export of heterologous passenger domains, opening up an effective tool for surface display of heterologous proteins. Here we report on the autotransporter domain of the Escherichia coli adhesin involved in diffuse adherence (AIDA-I), which was genetically fused to the C terminus of the periplasmic enzyme ␤-lactamase, leading to efficient expression of the fusion protein in E. coli. The ␤-lactamase moiety of the fusion protein was presented on the bacterial surface in a stable manner, and the surface-located ␤-lactamase was shown to be enzymatically active. Enzymatic activity was completely removed by protease treatment, indicating that surface display of ␤-lactamase was almost quantitative. The periplasmic domain of the outer membrane protein OmpA was not affected by externally added proteases, demonstrating that the outer membranes of E. coli cells expressing the ␤-lactamase AIDA-I fusion protein remained physiologically intact.

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Absence of periplasmic DsbA oxidoreductase facilitates export of cysteine-containing passenger proteins to the Escherichia coli cell surface via the Igaβ autotransporter pathway

Cited by 93 publications

References 10 publications

Type V Protein Secretion Pathway: the Autotransporter Story

Type V Protein Secretion Pathway: the Autotransporter Story

Autotransported Serine Protease A of Neisseria meningitidis : an Immunogenic, Surface-Exposed Outer Membrane, and Secreted Protein

Autodisplay: Functional Display of Active β-Lactamase on the Surface of Escherichia coli by the AIDA-I Autotransporter

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