Porcine ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (NPP1/CD203a): cloning, transcription, expression, mapping, and identification of an NPP1/CD203a epitope for swine workshop cluster 9 (SWC9) monoclonal antibodies.
Swine workshop cluster 9 (SWC9) antibody identifying a porcine epitope on macrophages and thymocytes was used to precipitate and characterize the molecule from biotinylated macrophages and to obtain peptide sequence by mass spectrometry. The protein was identified as ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (NPP1/CD203a). The porcine NPP1/CD203a encoding gene was mapped to chromosome 1 using a radiation hybrid panel, and transcription was investigated by RT-PCR analysis of several tissues. The cDNA was cloned and introduced into COS7 cells resulting in expression of functionally active enzyme and verification of the specificity of an SWC9 reacting monoclonal antibody. The antibody was used for immunohistochemical examination of various porcine tissues. Most prominent expression of NPP1/CD203a was found in lung macrophages and liver sinusoids.
Petersen
C B
CB
Department of Veterinary Pathobiology, The Royal Veterinary and Agricultural University, DK-1870 Frederiksberg, Denmark.
Nygård
A-B
AB
Viuff
B
B
Fredholm
M
M
Aasted
B
B
Salomonsen
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D018985:Epitopes, B-Lymphocyte / Q000235:genetics*; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D008099:Liver / Q000276:immunology / Q000378:metabolism; D008168:Lung / Q000276:immunology / Q000378:metabolism; D008264:Macrophages / Q000276:immunology / Q000378:metabolism*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010727:Phosphoric Diester Hydrolases / Q000235:genetics* / Q000378:metabolism*; D011755:Pyrophosphatases / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013552:Swine / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic
D000911:Antibodies, Monoclonal; D018985:Epitopes, B-Lymphocyte; D010727:Phosphoric Diester Hydrolases; C065962:ectonucleotide pyrophosphatase phosphodiesterase 1; D011755:Pyrophosphatases