title: MAP1272c encodes an NlpC/P60 protein, an antigen detected in cattle with Johne's disease.
abstract: The protein encoded by MAP1272c has been shown to be an antigen of Mycobacterium avium subsp. paratuberculosis that contains an NlpC/P60 superfamily domain found in lipoproteins or integral membrane proteins. Proteins containing this domain have diverse enzymatic functions that include peptidases, amidases, and acetyltransferases. The NlpC protein was examined in comparison to over 100 recombinant proteins and showed the strongest antigenicity when analyzed with sera from cattle with Johne's disease. To further localize the immunogenicity of NlpC, recombinant proteins representing defined regions were expressed and evaluated with sera from cattle with Johne's disease. The region from amino acids 74 to 279 was shown to be the most immunogenic. This fragment was also evaluated against a commercially available enzyme-linked immunosorbent assay (ELISA). Two monoclonal antibodies were produced in mice immunized with the full-length protein, and each recognized a distinct epitope. These antibodies cross-reacted with proteins from other mycobacterial species and demonstrated variable sizes of the proteins expressed from these subspecies. Both antibodies were further analyzed, and their interaction with MAP1272c and MAP1204 was characterized by a solution-based, luminescent binding assay. These tools provide additional means to study a strong antigen of M. avium subsp. paratuberculosis.
authors.affiliation: USDA-ARS-National Animal Disease Center, Ames, Iowa, USA. john.bannantine@ars.usda.gov
authors.forename: John P
authors.initials: JP
authors.lastname: Bannantine
authors.forename: Cari K
authors.initials: CK
authors.lastname: Lingle
authors.forename: Judith R
authors.initials: JR
authors.lastname: Stabel
authors.forename: Kasra X
authors.initials: KX
authors.lastname: Ramyar
authors.forename: Brandon L
authors.initials: BL
authors.lastname: Garcia
authors.forename: Alex J
authors.initials: AJ
authors.lastname: Raeber
authors.forename: Pascal
authors.initials: P
authors.lastname: Schacher
authors.forename: Vivek
authors.initials: V
authors.lastname: Kapur
authors.forename: Brian V
authors.initials: BV
authors.lastname: Geisbrecht
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000911:Antibodies, Monoclonal / Q000276:immunology; D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology*; D002417:Cattle; D002418:Cattle Diseases / Q000276:immunology / Q000382:microbiology; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D008074:Lipoproteins / Q000235:genetics / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016927:Mycobacterium avium subsp. paratuberculosis / Q000235:genetics* / Q000276:immunology*; D010283:Paratuberculosis / Q000276:immunology* / Q000382:microbiology*
chemical_list: D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D008074:Lipoproteins