Epstein E H, Bonifas J M
Hum Genet. 1985;71(3):201-5. doi: 10.1007/BF00284573.
Patients with recessive X-linked ichthyosis (RXLI), one hereditary form of scaly skin, lack activity of the enzyme steroid sulfatase in all tissues studied. To investigate the molecular defect underlying the lack of enzyme activity, we prepared antisera against normal enzyme by injecting normal placental microsomal suspensions or partially purified steroid sulfatase into rabbits. Antibody activity was assessed by immunoprecipitation of detergent solubilized steroid sulfatase. In addition, we prepared rabbit antisera against RXLI placental microsomal suspensions. To detect immunologically cross-reactive material in patients' placentas, extracts were studied by immunoblot techniques and by competition with normal enzyme for antibody binding. Patients' extracts did not contain immunoreactive material co-migrating on electrophoresis with purified enzyme nor did they inhibit immunoprecipitation of normal enzyme. Sera from rabbits immunized with RXLI placental microsomes contain no antibodies to normal steroid sulfatase, as judged by their failure to immunoprecipitate normal enzyme or to react with normal steroid sulfatase on immunoblot. Thus the mutation in RXLI appears to reduce steroid sulfatase enzyme protein as well as enzyme activity.
隐性X连锁鱼鳞病(RXLI)患者,一种遗传性鳞状皮肤病,在所有研究的组织中都缺乏类固醇硫酸酯酶的活性。为了研究酶活性缺乏背后的分子缺陷,我们通过将正常胎盘微粒体悬浮液或部分纯化的类固醇硫酸酯酶注射到兔子体内,制备了针对正常酶的抗血清。通过对去污剂溶解的类固醇硫酸酯酶进行免疫沉淀来评估抗体活性。此外,我们制备了针对RXLI胎盘微粒体悬浮液的兔抗血清。为了检测患者胎盘中的免疫交叉反应物质,通过免疫印迹技术以及与正常酶竞争抗体结合来研究提取物。患者的提取物在电泳时没有与纯化酶共迁移的免疫反应性物质,也没有抑制正常酶的免疫沉淀。用RXLI胎盘微粒体免疫的兔子血清中没有针对正常类固醇硫酸酯酶的抗体,这可以通过它们不能免疫沉淀正常酶或在免疫印迹上与正常类固醇硫酸酯酶反应来判断。因此,RXLI中的突变似乎降低了类固醇硫酸酯酶的酶蛋白以及酶活性。