Developmentally regulated localization and phosphorylation of SmIrV1, a Schistosoma mansoni antigen with similarity to calnexin.
Potential molecular targets of a protective humoral immune response against schistosomiasis have previously been identified based on their enhanced immunogenicity in mice vaccinated with irradiated cercaria as compared to chronically infected mice. One of these antigens, IrV1, has been molecularly cloned and its sequence shown to be similar to the molecular chaperone calnexin. In this investigation, we partially characterized IrV1 from different developmental stages of the schistosome. Immunoprecipitation studies with antibodies raised against a portion of recombinant IrV1 demonstrated its presence in cercaria, schistosomula, and adult worms with an apparent molecular mass on SDS-polyacrylamide gel electrophoresis of 90 kDa. There was an approximate 6-fold increase in protein expression level during the cercaria to schistosomula transformation. Consistent with a potential role as a molecular chaperone, IrV1 was associated with several metabolically labeled proteins in co-immunoprecipitation studies with the adult worm tegumental fraction. Similar to calnexin, IrV1 was metabolically labeled with 32P in adult worms on serine and threonine residues and was one of the major phosphoproteins of this stage. This phosphorylation was developmentally regulated and coincided with the transformation of cercaria into schistosomula. The localization was also stage-specific as IrV1 was transported from internal regions of cercaria to the outer tegumental layer of schistosomula. The presence of IrV1 on the surface of schistosomula, an unprecedented localization for this family of endoplasmic reticulum proteins, supports additional studies of the immunoprophylactic potential of this molecule.
Hawn
T R
TR
Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Strand
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis; D000913:Antibodies, Protozoan; D000947:Antigens, Helminth / Q000737:chemistry / Q000378:metabolism*; D000953:Antigens, Protozoan; D002135:Calcium-Binding Proteins / Q000737:chemistry*; D037281:Calnexin; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000737:chemistry*; D010766:Phosphorylation; D016052:Protozoan Vaccines; D012550:Schistosoma mansoni / Q000254:growth & development / Q000276:immunology*; D014612:Vaccines / Q000737:chemistry / Q000276:immunology
D000909:Antibodies, Helminth; D000913:Antibodies, Protozoan; D000947:Antigens, Helminth; D000953:Antigens, Protozoan; D002135:Calcium-Binding Proteins; D010750:Phosphoproteins; D016052:Protozoan Vaccines; D014612:Vaccines; D037281:Calnexin