Kuroiwa K, Arai T, Okazaki H, Minota S, Tominaga S
Department of Biochemistry, Jichi Medical School, Tochigi, 329-0498, Japan.
Biochem Biophys Res Commun. 2001 Jun 29;284(5):1104-8. doi: 10.1006/bbrc.2001.5090.
Soluble human ST2 protein (IL1RL1-a) in the sera of patients with various autoimmune diseases was identified by a newly developed procedure using specific monoclonal antibodies. After immunoprecipitation and subsequent immunoblotting, a glycosylated protein of about 60 kDa was detected in the sera of SLE patients, but not in the sera of healthy controls. The experiments using gel filtration and SDS-PAGE under a nonreducing condition indicated the existence of the ST2 multimer in serum. The mobility of the natural protein was slower than that of the recombinant human ST2 protein produced by COS7 cells in SDS-PAGE, suggesting a difference of glycosylation between humans and monkeys. The identification of the natural human ST2 protein should be important both to fundamental researches and the further clarification of the clinical implications of the ST2 protein.
采用新开发的使用特异性单克隆抗体的方法,在多种自身免疫性疾病患者的血清中鉴定出可溶性人ST2蛋白(IL1RL1-a)。经免疫沉淀及后续免疫印迹后,在系统性红斑狼疮(SLE)患者血清中检测到一种约60 kDa的糖基化蛋白,而在健康对照者血清中未检测到。在非还原条件下使用凝胶过滤和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)的实验表明血清中存在ST2多聚体。在SDS-PAGE中,天然蛋白的迁移率比COS7细胞产生的重组人ST2蛋白慢,这表明人和猴之间糖基化存在差异。天然人ST2蛋白的鉴定对于基础研究以及进一步阐明ST2蛋白的临床意义均应具有重要意义。