Isolation and analysis of cDNA clones expressing human lupus La antigen.

Several cDNA clones of the La antigen recognized by certain lupus autoantibodies were isolated from lambda gt11 expression libraries made from human liver. Recombinant clones were used to hybrid-select HeLa cell mRNA that was subsequently translated in vitro into a single protein species that comigrated with HeLa cell La protein. The in vitro translated protein was reactive with anti-La patient sera and was identical to the authentic La protein by peptide mapping. By analyzing overlapping cDNA clones, we mapped an antigenic site of La protein at the terminal 12% of the carboxyl end of the molecule. Within this region we identified a unique decapeptide of high hydrophilicity that may constitute a La antigenic determinant. We further demonstrated that the La antigen expressed from the recombinant clones can be used in a definitive enzyme-linked assay (ELISA) for the classification of sera from patients with systemic lupus erythematosus.

Duke Scholars

Author Jack Donald Keene Molecular Genetics and Microbiology