FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2896724204> ?p ?o ?g. }

• W2896724204 abstract "Plague is a notifiable disease caused by Yersinia pestis for which there is still no reliablevaccine available. The fraction 1 (F1) antigen forms an immunogenic capsule around the cellsurface that decreases efficiency of phagocytosis. Many E. coli strains and other Gramnegativebacteria coat themselves in similar, remarkably stable protein fibres assembled viaspecific, dedicated chaperone-usher (CU) pathways. Fibres are assembled through a porewithin each usher protein, via a cycling of specific chaperone:subunit:usher interactionsleading to subunit polymerisation and translocation to the cell surface. This project has usedbioinformatics analysis and modelling to improve understanding of details of the Y. pestis F1chaperone:usher translocon and has applied this knowledge to enhance export ofheterologous epitopes. The number of members of the γ3 family of ushers, which assemblesimple fibres of one or two subunits, has been expanded and includes for the first time a CUsystem more closely related to that of the Yersinia Caf system. This plasmid encoded usherfrom the commensal E. coli SE11 shares 70% identity with Caf1A. Caf1A usher wasmodelled using Intfold and ITASSER, based on the crystal structures of E. coli FimD andPapC usher in the closed state (PDB: 2VQI, 3.2A). The complete Caf translocon wasmodelled based on the open FimD structure (PDB: 3RFZ, 2.8A) with the Caf1Mchaperone:Caf1:Caf1 subunit structure (PDB:1Z9S, 2.2A) docked in the translocon. Basedon the modelled Caf translocon and multiple sequence alignments of related ushers,conserved residues within the β-barrel facing the pore cavity or subunit; residues interactingbetween the plug and β-barrel in the closed pore and residues interacting between β-barreland subunit in the ‘open’ translocating pores were identified. These were then tested bymutagenesis for impact on F1 assembly. No single point mutation within the usher abolishedF1 assembly. The most profound effect was following mutation of Ser289 to Ala, the levelof surface F1 decreased to 70.81% ± 3.0 of levels with wildtype Caf1A. The potential of F1fibre to act as a carrier of epitopes was tested by replacing 4 loops within Caf1 with eitherGly residues or charged residues. The model was used to understand and optimise limitationsin export of modified fibres. One permissive site (loop 5) was identified as the optimum sitefor loop replacement and surface assembly of modified F1 was enhanced by mutation of aclashing Gln in the β-barrel wall. In addition, the study identified a critical role for aconserved Asn, Asn80, within loop 2 of Caf1 subunit that was modelled interacting withTyr266 in the plug. The permissive loop 5 was also adapted to export a polyhistidinesequence. While 4 His residues were readily accommodated, F1::1His6 conferred toxicity,although there was still evidence of surface polymer. Thus, this study demonstrated flexibilityof the F1 CU pathway for surface display of short peptides and the ability to use thetranslocon model to identify problem areas and repair export. As F1 is considered to beunique to Y. pestis, the presence of a Caf related CU locus in a commensal E. coli was ofinterest to understanding its phylogenetic relationship to F1 and also in diagnosticapplications in plague detection. Bioinformatic analysis and recombinant expression of theE. coli SE11-P2 CU locus, combined with mass spectrometry analysis, confirmed surfaceassembly of polymers of both subunits from this locus. A purification strategy for isolationof recombinant SE11-P2 polymers was developed. Purified protein will be used in futurestudies to raise antibody for further study of this CU system and to test for any cross-reactionwith Y. pestis F1 antigen, that might potentially interfere with serological tests for Y. pestis.F1 antigen remains a component of interest in both anti-plague vaccine design and plaguedetection. Results from this study have potential to improve reliability of both approaches toplague control." @default.
• W2896724204 created "2018-10-26" @default.
• W2896724204 creator A5084643603 @default.
• W2896724204 date "2017-01-01" @default.
• W2896724204 modified "2023-09-23" @default.
• W2896724204 title "The F1 chaperone:usher translocon of Yersinia pestis and potential applications" @default.
• W2896724204 hasPublicationYear "2017" @default.
• W2896724204 type Work @default.
• W2896724204 sameAs 2896724204 @default.
• W2896724204 citedByCount "0" @default.
• W2896724204 crossrefType "dissertation" @default.
• W2896724204 hasAuthorship W2896724204A5084643603 @default.
• W2896724204 hasConcept C104292427 @default.
• W2896724204 hasConcept C104317684 @default.
• W2896724204 hasConcept C142724271 @default.
• W2896724204 hasConcept C144647389 @default.
• W2896724204 hasConcept C199229279 @default.
• W2896724204 hasConcept C2775962898 @default.
• W2896724204 hasConcept C2778222588 @default.
• W2896724204 hasConcept C41625074 @default.
• W2896724204 hasConcept C55493867 @default.
• W2896724204 hasConcept C60987743 @default.
• W2896724204 hasConcept C65556437 @default.
• W2896724204 hasConcept C71924100 @default.
• W2896724204 hasConcept C86803240 @default.
• W2896724204 hasConcept C95444343 @default.
• W2896724204 hasConceptScore W2896724204C104292427 @default.
• W2896724204 hasConceptScore W2896724204C104317684 @default.
• W2896724204 hasConceptScore W2896724204C142724271 @default.
• W2896724204 hasConceptScore W2896724204C144647389 @default.
• W2896724204 hasConceptScore W2896724204C199229279 @default.
• W2896724204 hasConceptScore W2896724204C2775962898 @default.
• W2896724204 hasConceptScore W2896724204C2778222588 @default.
• W2896724204 hasConceptScore W2896724204C41625074 @default.
• W2896724204 hasConceptScore W2896724204C55493867 @default.
• W2896724204 hasConceptScore W2896724204C60987743 @default.
• W2896724204 hasConceptScore W2896724204C65556437 @default.
• W2896724204 hasConceptScore W2896724204C71924100 @default.
• W2896724204 hasConceptScore W2896724204C86803240 @default.
• W2896724204 hasConceptScore W2896724204C95444343 @default.
• W2896724204 hasLocation W28967242041 @default.
• W2896724204 hasOpenAccess W2896724204 @default.
• W2896724204 hasPrimaryLocation W28967242041 @default.
• W2896724204 hasRelatedWork W1964303928 @default.
• W2896724204 hasRelatedWork W1979325155 @default.
• W2896724204 hasRelatedWork W1986789701 @default.
• W2896724204 hasRelatedWork W2012123122 @default.
• W2896724204 hasRelatedWork W2019203602 @default.
• W2896724204 hasRelatedWork W2026957483 @default.
• W2896724204 hasRelatedWork W2033947606 @default.
• W2896724204 hasRelatedWork W2042588480 @default.
• W2896724204 hasRelatedWork W2050702257 @default.
• W2896724204 hasRelatedWork W2059486580 @default.
• W2896724204 hasRelatedWork W2081523176 @default.
• W2896724204 hasRelatedWork W2091115293 @default.
• W2896724204 hasRelatedWork W2094502729 @default.
• W2896724204 hasRelatedWork W2107851838 @default.
• W2896724204 hasRelatedWork W2112638132 @default.
• W2896724204 hasRelatedWork W2118044534 @default.
• W2896724204 hasRelatedWork W2127241473 @default.
• W2896724204 hasRelatedWork W2145409351 @default.
• W2896724204 hasRelatedWork W2151763805 @default.
• W2896724204 hasRelatedWork W96349316 @default.
• W2896724204 isParatext "false" @default.
• W2896724204 isRetracted "false" @default.
• W2896724204 magId "2896724204" @default.
• W2896724204 workType "dissertation" @default.