Nucleotide and amino acid sequences and immunochemical characterization of the C-terminal region of Nedd5. (a) The nucleotide sequence of 335 base pairs of the cloned cDNA insertion was sequenced with automated sequencer ABI Prism 310 Collection. (b) The amino acid sequence predicted from the nucleotide sequence is 109 residues long. The sequence compared with the GenBank sequence database using both Fasta and Blast analysis has 100% identity with the C-terminal subunit of Nedd5 (accession number Q15019). (c) The molecular size and the purity of the expressed protein were confirmed by 12% SDS–PAGE stained by Coomassie blue (lane 1). Immunoreactivity was analyzed by immunoblotting: monoclonal antibody antihistidine tail (lane 2); mouse polyclonal antiserum specific to Nedd5 C-ter (lane 3); representative sera from three patients with SLE IgG positive to Nedd5 (lanes 4,5,6); representative serum from a patient with SLE IgG negative to Nedd5 C-ter (lane 7); representative serum from a healthy subject (lane 8); control lane without serum (lane 9).