Pacheco Benny, Malmström Anders, Maccarana Marco
Department of Experimental Medical Science, Biomedical Center D12, Lund University, Tornavägen 10, SE-221 84 Lund, Sweden.
J Biol Chem. 2009 Apr 10;284(15):9788-95. doi: 10.1074/jbc.M809339200. Epub 2009 Feb 2.
A second dermatan sulfate epimerase (DS-epi2) was identified as a homolog of the first epimerase (DS-epi1), which was previously described by our group. DS-epi2 is 1,222 amino acids long and has an approximately 700-amino acid N-terminal epimerase domain that is highly conserved between the two enzymes. In addition, the C-terminal portion is predicted to be an O-sulfotransferase domain. In this study we found that DS-epi2 has epimerase activity, which involves conversion of d-glucuronic acid to l-iduronic acid (EC 5.1.3.19), but no O-sulfotransferase activity was detected. In dermatan sulfate, iduronic acid residues are either clustered together in blocks or alternating with glucuronic acid, forming hybrid structures. By using a short interfering RNA approach, we found that DS-epi2 and DS-epi1 are both involved in the biosynthesis of the iduronic acid blocks in fibroblasts and that DS-epi2 can also synthesize the hybrid structures. Both iduronic acid-containing domains have been shown to bind to several growth factors, many of which have biological roles in brain development. DS-epi2 has been genetically linked to bipolar disorder, which suggests that the dermatan sulfate domains generated by a defective enzyme may be involved in the etiology of the disease.
第二种硫酸皮肤素表异构酶(DS-epi2)被鉴定为第一种表异构酶(DS-epi1)的同源物,我们团队之前已对DS-epi1进行过描述。DS-epi2由1222个氨基酸组成,其N端约700个氨基酸的表异构酶结构域在这两种酶之间高度保守。此外,预测其C端部分为O-磺基转移酶结构域。在本研究中,我们发现DS-epi2具有表异构酶活性,可将D-葡糖醛酸转化为L-艾杜糖醛酸(EC 5.1.3.19),但未检测到O-磺基转移酶活性。在硫酸皮肤素中,艾杜糖醛酸残基要么成簇聚集在一起,要么与葡糖醛酸交替出现,形成杂合结构。通过使用短干扰RNA方法,我们发现DS-epi2和DS-epi1都参与成纤维细胞中艾杜糖醛酸块的生物合成,并且DS-epi2还能合成杂合结构。含艾杜糖醛酸的结构域均已被证明可与多种生长因子结合,其中许多生长因子在大脑发育中具有生物学作用。DS-epi2在基因上与双相情感障碍有关,这表明由缺陷酶产生的硫酸皮肤素结构域可能参与了该疾病的病因。