Common structural features of IgA1 protease‐like outer membrane protein autotransporters

Jose, Joachim; Jähnig, Fritz; Meyer, Thomas F.

doi:10.1111/j.1365-2958.1995.mmi_18020378.x

Cited by 150 publications

(137 citation statements)

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“…All of these proteins are translated as large precursor proteins that are capable of translocation across the outer membrane, apparently in the absence of accessory factors or energy coupling. In each case, the C-terminus is predicted to form an outer membrane ␤-barrel with amphipathic, antiparallel ␤-strands that generate a hydrophobic coat in contact with the membrane and a hydrophilic interior (Jose et al, 1995). The ␤-barrel pore is presumed to serve as a conduit for extrusion of the N-terminal passenger domain, allowing for surface localization.…”

Section: Discussionmentioning

confidence: 99%

“…Comparison of Hap ␤ with outer membrane proteins that form ␤-barrel structures revealed the presence of a consensus amino acid motif at the C-terminus (Struvyè et al, 1991;Jose et al, 1995). In these proteins, the last ᮊ 1997 Blackwell Science Ltd, Molecular Microbiology, 26, 505-518 Schirmer and Cowan (1993).…”

Section: Hap Is a Serine Protease And Undergoes Autoproteolytic Cleavagementioning

confidence: 99%

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Structural determinants of processing and secretion of the Haemophilus influenzae Hap protein

Hendrixson

Morena

Stathopoulos

et al. 1997

Molecular Microbiology

102

138

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SummaryHaemophilus influenzae elaborates a surface protein called Hap, which is associated with the capacity for intimate interaction with cultured epithelial cells. Expression of hap results in the production of three protein species: outer membrane proteins of approximately 155 kDa and 45 kDa and an extracellular protein of approximately 110 kDa. The 155 kDa protein corresponds to full-length mature Hap (without the signal sequence), and the 110 kDa extracellular protein represents the N-terminal portion of mature Hap (designated Hap s ). In the present study, we examined the mechanism of processing and secretion of Hap. Site-directed mutagenesis suggested that Hap is a serine protease that undergoes autoproteolytic cleavage to generate the 110 kDa extracellular protein and the 45 kDa outer membrane protein. Biochemical analysis confirmed this conclusion and established that cleavage occurs on the bacterial cell surface. Determination of N-terminal amino acid sequence and mutagenesis studies revealed that the 45 kDa protein corresponds to the Cterminal portion of Hap, starting at N1037. Analysis of the secondary structure of this protein (designated Hap ␤ ) predicted formation of a ␤-barrel with an N-terminal transmembrane ␣-helix followed by 14 transmembrane ␤-strands. Additional analysis revealed that the final ␤-strand contains an amino acid motif common to other ␤-barrel outer membrane proteins. Upon deletion of this entire C-terminal consensus motif, Hap could no longer be detected in the outer membrane, and secretion of Hap s was abolished. Deletion or complete alteration of the final three amino acid residues had a similar but less dramatic effect, suggesting that this terminal tripeptide is particularly important for outer membrane localization and/or stability of the protein. In contrast, isolated point mutations that disrupted the amphipathic nature of the consensus motif or eliminated the C-terminal tryptophan had no effect on outer membrane localization of Hap or secretion of Hap s . These results provide insight into a growing family of Gram-negative bacterial exoproteins that are secreted by an IgA1 protease-like mechanism; in addition, they contribute to a better understanding of the structural determinants of targeting of ␤-barrel proteins to the bacterial outer membrane.

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Section: Discussionmentioning

confidence: 99%

Section: Hap Is a Serine Protease And Undergoes Autoproteolytic Cleavagementioning

confidence: 99%

Structural determinants of processing and secretion of the Haemophilus influenzae Hap protein

Hendrixson

Morena

Stathopoulos

et al. 1997

Molecular Microbiology

102

138

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“…Identified meningococcal putative proteins were analyzed for the characteristic features exhibited by autotransporter proteins including: size (Ͼ60 kDa); a low cysteine content (two to five); and the presence of a signal sequence and a C-terminal consensus sequence, (Y/V/I/F/W)-x-(F/W) (15,17). The search and analysis identified a previously uncharacterized CDS (NMA0478) with a deduced molecular mass of ca.…”

Section: Identification Of a Novel Meningococcal Autotransporter Protmentioning

confidence: 99%

“…The autotransporter mechanism of protein transportation and secretion was first described for the immunoglobulin A1 (IgA1) proteases of pathogenic Neisseria (26) and is a rapidly expanding family of proteins (14,15). The characteristic structure of autotransporter proteins and the secretion mechanism have been reviewed elsewhere (15,17). Autotransporter proteins, possessing a diverse array of N-terminal "functional" domains, have been reported in many gram-negative organisms (1,2,5,6,9,10,12,13,(23)(24)(25)(29)(30)(31).…”

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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 secretion systems are classified into five types based on function and sequence (1)(2)(3)(4)(5)(6). The type I, II, III, and IV systems are unified by the presence of at least one potential NTP hydrolase (NTPase) protein.…”

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confidence: 99%

Phylogeny of genes for secretion NTPases: Identification of the widespread tadA subfamily and development of a diagnostic key for gene classification

Planet

Kachlany

DeSalle

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

188

166

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Macromolecular transport systems in bacteria currently are classified by function and sequence comparisons into five basic types. In this classification system, type II and type IV secretion systems both possess members of a superfamily of genes for putative NTP hydrolase (NTPase) proteins that are strikingly similar in structure, function, and sequence. These include VirB11, TrbB, TraG, GspE, PilB, PilT, and ComG1. The predicted protein product of tadA, a recently discovered gene required for tenacious adherence of Actinobacillus actinomycetemcomitans, also has significant sequence similarity to members of this superfamily and to several unclassified and uncharacterized gene products of both Archaea and Bacteria. To understand the relationship of tadA and tadA-like genes to those encoding the putative NTPases of type II͞IV secretion, we used a phylogenetic approach to obtain a genealogy of 148 NTPase genes and reconstruct a scenario of gene superfamily evolution. In this phylogeny, clear distinctions can be made between type II and type IV families and their constituent subfamilies. In addition, the subgroup containing tadA constitutes a novel and extremely widespread subfamily of the family encompassing all putative NTPases of type IV secretion systems. We report diagnostic amino acid residue positions for each major monophyletic family and subfamily in the phylogenetic tree, and we propose an easy method for precisely classifying and naming putative NTPase genes based on phylogeny. This molecular key-based method can be applied to other gene superfamilies and represents a valuable tool for genome analysis.conjugation ͉ ATPase ͉ Archaea ͉ molecular key ͉ Walker box

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Common structural features of IgA1 protease‐like outer membrane protein autotransporters

Cited by 150 publications

References 0 publications

Structural determinants of processing and secretion of the Haemophilus influenzae Hap protein

Structural determinants of processing and secretion of the Haemophilus influenzae Hap protein

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

Phylogeny of genes for secretion NTPases: Identification of the widespread tadA subfamily and development of a diagnostic key for gene classification

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