title: Fructose 1,6-Bisphosphate Aldolase, a Novel Immunogenic Surface Protein on Listeria Species.
abstract: Listeria monocytogenes is a ubiquitous food-borne pathogen, and its presence in food or production facilities highlights the importance of surveillance. Increased understanding of the surface exposed antigens on Listeria would provide potential diagnostic and therapeutic targets. In the present work, using mass spectrometry and genetic cloning, we show that fructose-1,6-bisphosphate aldolase (FBA) class II in Listeria species is the antigen target of the previously described mAb-3F8. Western and dot blot assays confirmed that the mAb-3F8 could distinguish all tested Listeria species from close-related bacteria. Localization studies indicated that FBA is present in every fraction of Listeria cells, including supernatant and the cell wall, setting Listeria spp. as one of the few bacteria described to have this protein on their cell surface. Epitope mapping using ORFeome display and a peptide membrane revealed a 14-amino acid peptide as the potential mAb-3F8 epitope. The target epitope in FBA allowed distinguishing Listeria spp. from closely-related bacteria, and was identified as part of the active site in the dimeric enzyme. However, its function in cell surface seems not to be host cell adhesion-related. Western and dot blot assays further demonstrated that mAb-3F8 together with anti-InlA mAb-2D12 could differentiate pathogenic from non-pathogenic Listeria isolated from artificially contaminated cheese. In summary, we report FBA as a novel immunogenic surface target useful for the detection of Listeria genus.
authors.affiliation: Laboratório de Imunologia Aplicada, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.|Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, Indiana, United Sates of America.
authors.forename: Marcelo
authors.initials: M
authors.lastname: Mendonça
authors.affiliation: Laboratório de Imunologia Aplicada, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.|Technische Universität Braunschweig, Institut für Biochemie, Biotechnologie und Bioinformatik, Abteilung Biotechnologie, Spielmannstr, Braunschweig, Germany.
authors.forename: Gustavo Marçal Schmidt Garcia
authors.initials: GM
authors.lastname: Moreira
authors.affiliation: Laboratório de Imunologia Aplicada, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Fabricio Rochedo
authors.initials: FR
authors.lastname: Conceição
authors.affiliation: Technische Universität Braunschweig, Institut für Biochemie, Biotechnologie und Bioinformatik, Abteilung Biotechnologie, Spielmannstr, Braunschweig, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Hust
authors.affiliation: Laboratório de Microbiologia de Alimentos, Departamento de Ciência e Tecnologia Agroindustrial, Faculdade de Agronomia Eliseu Maciel, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Karla Sequeira
authors.initials: KS
authors.lastname: Mendonça
authors.affiliation: Laboratório de Imunologia Aplicada, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Ângela Nunes
authors.initials: ÂN
authors.lastname: Moreira
authors.affiliation: Laboratório de Imunologia Aplicada, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Rodrigo Correa
authors.initials: RC
authors.lastname: França
authors.affiliation: Laboratório de Microbiologia de Alimentos, Departamento de Ciência e Tecnologia Agroindustrial, Faculdade de Agronomia Eliseu Maciel, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Wladimir Padilha
authors.initials: WP
authors.lastname: da Silva
authors.affiliation: Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, Indiana, United Sates of America.
authors.forename: Arun K
authors.identifier: http://orcid.org/0000-0003-3640-1554
authors.initials: AK
authors.lastname: Bhunia
authors.affiliation: Laboratório de Imunologia Aplicada, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: José Antonio G
authors.initials: JA
authors.lastname: Aleixo
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D000954:Antigens, Surface / Q000276:immunology* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015153:Blotting, Western; D020134:Catalytic Domain; D002611:Cheese / Q000382:microbiology; D003001:Cloning, Molecular; D019281:Dimerization; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology; D005516:Food Microbiology; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D008087:Listeria / Q000201:enzymology* / Q000276:immunology* / Q000302:isolation & purification; D013058:Mass Spectrometry; D010455:Peptides / Q000032:analysis; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D001426:Bacterial Proteins; D000939:Epitopes; D010455:Peptides; D011994:Recombinant Proteins; D005634:Fructose-Bisphosphate Aldolase