Damage-induced neuronal endopeptidase (DINE) is a unique metallopeptidase expressed in response to neuronal damage and activates superoxide scavengers -- Data Supplement - Supplemental Fig. 5 -- Proceedings of the National Academy of Sciences

Supplementary material for Kiryu-Seo et al., Proc. Natl. Acad. Sci. USA, 10.1073/pnas.070509897

Fig. 5.
Structure of DINE. (A) Alignment of alternatively spliced forms of transcripts. Bars represent probes (probes a and b) used for Northern blotting and RNase protection assay. A mouse genome library (lFIXII Library, Stratagene) was screened with a subcloned PCR-derived DNA fragment to obtain mouse DINE genome. Five positive plaques were obtained, purified, and partially sequenced. By using rat genome extracted from liver, PCR with LA taq polymerase was performed to obtain rat DINE genome near the stop codon. The PCR fragment was subcloned in pGEM-T vector (Promega) and sequenced. The genome structure of 3' side of ORF and 3' noncoding region is magnified below. Closed boxes denote exons and arrow shows stop codon. (B) Rat and mouse DINE amino acids sequence. The predicted transmembrane domain is underlined. A zinc-binding motif is surrounded by a box. The asterisk shows conserved amino acid among NEP family. The nucleotide sequences reported in this paper have been deposited in the GenBank database (accession nos. AB023896, AB026293, and AB026294). (C) Tissue localization of DINE mRNA by RNase protection assay using probe b and a probe for glyceraldehyde-3-phosphate dehydrogenase (GAPDH).