We identified the gene encoding
chondroitin-
glucuronate C5-epimerase (EC 5.1.3.19) that converts D-
glucuronic acid to L-
iduronic acid residues in
dermatan sulfate biosynthesis. The
enzyme was solubilized from bovine spleen, and an approximately 43,000-fold purified preparation containing a major 89-kDa candidate component was subjected to mass spectrometry analysis of tryptic
peptides. SART2 (
squamous cell carcinoma antigen recognized by T cell 2), a
protein with unknown function highly expressed in
cancer cells and tissues, was identified by 18
peptides covering 26% of the sequence. Transient expression of
cDNA resulted in a 22-fold increase in
epimerase activity in 293HEK cell lysate. Moreover, overexpressing cells produced
dermatan sulfate chains with 20% of
iduronic acid-containing
disaccharide units, as compared with 5% for mock-transfected cells. The
iduronic acid residues were preferentially clustered in blocks, as in naturally occurring
dermatan sulfate. Given the discovered identity, we propose to rename SART2 (Nakao, M., Shichijo, S., Imaizumi, T., Inoue, Y., Matsunaga, K., Yamada, A., Kikuchi, M., Tsuda, N., Ohta, K., Takamori, S., Yamana, H., Fujita, H., and Itoh, K. (2000) J. Immunol. 164, 2565-2574) with a functional designation,
chondroitin-
glucuronate C5-epimerase (or DS
epimerase). DS
epimerase activity is ubiquitously present in normal tissues, although with marked quantitative differences. It is highly homologous to part of the NCAG1
protein, encoded by the C18orf4 gene, genetically linked to
bipolar disorder. NCAG1 also contains a putative
chondroitin sulfate sulfotransferase domain and thus may be involved in
dermatan sulfate biosynthesis. The functional relation between
dermatan sulfate and
cancer is unknown but may involve known
iduronic acid-dependent interactions with
growth factors,
selectins,
cytokines, or coagulation inhibitors.