Sequence analysis and recombinant expression of a 28-kilodalton Treponema pallidum subsp. pallidum rare outer membrane protein (Tromp2)

Champion, Cheryl I.; Blanco, David R.; Exner, Maurice M.; Erdjument‐Bromage, Hediye; Hancock, Robert E. W.; Tempst, Paul; Miller, James N.; Lovett, Michael

doi:10.1128/jb.179.4.1230-1238.1997

Cited by 19 publications

(12 citation statements)

References 36 publications

(40 reference statements)

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“…Definition of the mechanism of immunity and the location of immunoprotective epitopes could also make it possible to use systems for recombinant protein expression which are more efficient than membrane protein expression. The results of these studies should have relevance to vaccine development in spirochetal and other bacterial diseases in which OMPs are being explored as potential vaccinogens (7,14,24,25,52,53).…”

Section: Discussionmentioning

confidence: 99%

Leptospiral Outer Membrane Proteins OmpL1 and LipL41 Exhibit Synergistic Immunoprotection

et al. 1999

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New vaccine strategies are needed for prevention of leptospirosis, a widespread human and veterinary disease caused by invasive spirochetes belonging to the genus Leptospira. We have examined the immunoprotective capacity of the leptospiral porin OmpL1 and the leptospiral outer membrane lipoprotein LipL41 in the Golden Syrian hamster model of leptospirosis. Specialized expression plasmids were developed to facilitate expression of leptospiral proteins inEscherichia coli as the membrane-associated proteins OmpL1-M and LipL41-M. Although OmpL1-M expression is highly toxic inE. coli, this was accomplished by using plasmid pMMB66-OmpL1, which has undetectable background expression without induction. LipL41-M expression and processing were enhanced by altering its lipoprotein signal peptidase cleavage site to mimic that of the murein lipoprotein. Active immunization of hamsters with E. coli membrane fractions containing a combination of OmpL1-M and LipL41-M was found to provide significant protection against homologous challenge with Leptospira kirschneri serovar grippotyphosa. At 28 days after intraperitoneal inoculation, survival in animals vaccinated with both proteins was 71% (95% confidence interval [CI], 53 to 89%), compared with only 25% (95% CI, 8 to 42%) in the control group (P < 0.001). On the basis of serological, histological, and microbiological assays, no evidence of infection was found in the vaccinated survivors. The protective effects of immunization with OmpL1-M and LipL41-M were synergistic, since significant levels of protection were not observed in animals immunized with either OmpL1-M or LipL41-M alone. In contrast to immunization with the membrane-associated forms of leptospiral proteins, hamsters immunized with His6-OmpL1 and His6-LipL41 fusion proteins, either alone or in combination, were not protected. These data indicate that the manner in which OmpL1 and LipL41 associates with membranes is an important determinant of immunoprotection.

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Section: Discussionmentioning

confidence: 99%

Leptospiral Outer Membrane Proteins OmpL1 and LipL41 Exhibit Synergistic Immunoprotection

et al. 1999

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“…These include characterized f lagellar components, FliL (TDE2764), FliE (TDE1214) (17), putative outer layer protein FlaA (TDE1408) (18) and its distant paralogs TDE1712 and TDE1409, that *The distribution of CDSs in the T. pallidum and B. burgdorferi chromosomes are derived from the original annotation. These numbers, particularly hypothetical and conserved hypothetical proteins, may be significantly different with updated blast searches and annotation.…”

Section: Resultsmentioning

confidence: 99%

Comparison of the genome of the oral pathogen Treponema denticola with other spirochete genomes

Seshadri

Myers

Tettelin

et al. 2004

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

253

314

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We present the complete 2,843,201-bp genome sequence of Treponema denticola (ATCC 35405) an oral spirochete associated with periodontal disease. Analysis of the T. denticola genome reveals factors mediating coaggregation, cell signaling, stress protection, and other competitive and cooperative measures, consistent with its pathogenic nature and lifestyle within the mixed-species environment of subgingival dental plaque. Comparisons with previously sequenced spirochete genomes revealed specific factors contributing to differences and similarities in spirochete physiology as well as pathogenic potential. The T. denticola genome is considerably larger in size than the genome of the related syphiliscausing spirochete Treponema pallidum. The differences in gene content appear to be attributable to a combination of three phenomena: genome reduction, lineage-specific expansions, and horizontal gene transfer. Genes lost due to reductive evolution appear to be largely involved in metabolism and transport, whereas some of the genes that have arisen due to lineage-specific expansions are implicated in various pathogenic interactions, and genes acquired via horizontal gene transfer are largely phagerelated or of unknown function.

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“…A major outcome of these endeavors has been the development of methods for fractionation of T. pallidum and isolation of OMs in quantities sufficient for microsequence analysis of protein constituents (5,9,35). At the same time, the availability of these methods has exposed our rather limited inventory of T. pallidum cell envelope constituents (29) and, more importantly, the dearth of monospecific immunologic reagents for identifying bona fide OM proteins.…”

Section: Discussionmentioning

confidence: 99%

Molecular characterization and cellular localization of TpLRR, a processed leucine-rich repeat protein of Treponema pallidum, the syphilis spirochete

et al. 1997

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Automated Edman degradation was used to obtain N-terminal and internal amino acid sequences from a 26-kDa protein in isolated Treponema pallidum outer membranes (OMs). The resulting sequences enabled us to PCR amplify from T. pallidum DNA a 275-bp fragment of the corresponding gene. The complete nucleotide sequence of the gene was determined from fragments amplified by long-distance PCR. Primer extension verified the assigned translational start of the open reading frame (ORF) and putative upstream promoter elements. The ORF encoded a highly basic (pI 9.6) 26-kDa protein which contained an N-terminal 25-aminoacid leader peptide terminated by a signal peptidase I cleavage site. The mature protein contained seven tandemly spaced copies (as well as an eighth incomplete copy) of a leucine-rich repeat (LRR), a motif previously identified in a number of prokaryotic and eukaryotic proteins. Accordingly, the polypeptide was designated T. pallidum leucine-rich repeat protein (TpLRR). Although Triton X-114 phase partitioning showed that TpLRR was hydrophilic, cell localization studies showed that most of the antigen was associated with the peptidoglycan-cytoplasmic membrane complex rather than being freely soluble in the periplasmic space. Immunoblot studies showed that syphilis patients develop a weak antibody response to the antigen. Lastly, the lrr T. pallidum gene was mapped to a 60-kb SfiI-SpeI fragment of the T. pallidum chromosome which also contains the rrnA and flaA genes. The function(s) of TpLRR is currently unknown; however, protein-protein and/or protein-lipid interactions mediated by its LRR motifs may facilitate interactions between components of the T. pallidum cell envelope.Syphilis is a sexually transmitted, multistage infection caused by the noncultivatable spirochetal pathogen Treponema pallidum (26). Like all spirochetes, T. pallidum is a highly motile bacterium in which an outer membrane (OM) surrounds the periplasmic space, peptidoglycan (PG)-cytoplasmic membrane (CM) complex, and protoplasmic cylinder (PC); within the periplasmic space are endoflagella, the organelles of motility (19). Compared with conventional gram-negative bacteria, we know comparatively little about the structure and composition of the T. pallidum cell envelope and how its diverse components promote survival of the syphilis spirochete in the demanding milieu of the mammalian host. Studies from a number of laboratories have provided compelling evidence that OMs of T. pallidum and gram-negative bacteria differ markedly with respect to physical properties, composition, and molecular architecture (6,29,32,34,42). While the paucity of surfaceexposed proteins appears to explain, at least in part, how this extracellular pathogen so successfully evades host immune responses during persistent infection, it also raises intriguing and fundamental questions regarding the mechanisms by which the bacterium acquires nutrients from its environment (32). In contrast to the protein-deficient OM, the vast majority of T. pallidum cell envelope con...

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Sequence analysis and recombinant expression of a 28-kilodalton Treponema pallidum subsp. pallidum rare outer membrane protein (Tromp2)

Cited by 19 publications

References 36 publications

Leptospiral Outer Membrane Proteins OmpL1 and LipL41 Exhibit Synergistic Immunoprotection

Leptospiral Outer Membrane Proteins OmpL1 and LipL41 Exhibit Synergistic Immunoprotection

Comparison of the genome of the oral pathogen Treponema denticola with other spirochete genomes

Molecular characterization and cellular localization of TpLRR, a processed leucine-rich repeat protein of Treponema pallidum, the syphilis spirochete

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