Chronic stress has been the popular phenomenon in the modern society. But we have not yet resolved the problem effectively. A novel protein in pig spleen, which possesses immunosuppressive activity, has been purified ...Chronic stress has been the popular phenomenon in the modern society. But we have not yet resolved the problem effectively. A novel protein in pig spleen, which possesses immunosuppressive activity, has been purified in this study. Its molecular weight about 200 kDa and three subunits composition were fractionated by SDS-PAGE. The protein inhibited mouse T-lymphocyte proliferation by the induced concanavalin A (ConA) and regulatory volume decrease (RVD), which may be due to the decreased Kv1.3 expression and activity. In addition, we have raised successfully its polyclonal antibody, which could reverse the inhibited lymphocyte proliferation by the immuno-suppressive protein. And the antibody could be used to coat plate for ELISA assay. Therefore, the antibody will have the potential to provide a basis for applying to clinical stress diseases in the future.展开更多
文摘Mink相关蛋白1(MiRP1)是由KCNE基因家族成员KCNE2编码的具有一个跨膜结构的小分子蛋白质,发生在KCNE2上的相关突变能够引起遗传性长QT间期综合症(long QT syndrome,LQT6),但其机制不明.以往的工作表明,MiRP1调节瞬间外向钾电流(transient outward current,Ito)的功能,对维持心电稳定性具有重要的调节作用.在哺乳细胞系COS-7表达系统利用膜片钳全细胞记录方式,研究了两种LQT6相关的突变体I57T和V65M对Kv4.3通道功能的影响,从MiRP1对Ito功能调控的改变探讨LQT6引起心律失常的电生理机制.结果表明,KCNE2与Kv4.3共表达后对通道功能具有明显的调控作用,使通道的激活和失活明显减慢,电压依赖性失活发生正向移位,同时加快Kv4.3通道从失活中的恢复.I57T与Kv4.3共表达的通道,门控动力学以及通道的恢复特性更接近Kv4.3单独表达的通道,表现为丧失KCNE2的功能——"loss of function",而V65M的作用则与之刚好相反,对Kv4.3门控动力学和恢复特性的调节较KCNE2更强,同时,使通道电流密度明显降低,表现为增强KCNE2的功能——"gain of function".由此推论,KCNE2对Ito功能有重要的调节作用,发生在KCNE2基因上的突变,无论是增强(V65M)还是减弱(I57T)KCNE2的功能都可能通过改变Ito在心脏电稳定性中的贡献,从而使心脏在某些条件下发生心律失常.
文摘Chronic stress has been the popular phenomenon in the modern society. But we have not yet resolved the problem effectively. A novel protein in pig spleen, which possesses immunosuppressive activity, has been purified in this study. Its molecular weight about 200 kDa and three subunits composition were fractionated by SDS-PAGE. The protein inhibited mouse T-lymphocyte proliferation by the induced concanavalin A (ConA) and regulatory volume decrease (RVD), which may be due to the decreased Kv1.3 expression and activity. In addition, we have raised successfully its polyclonal antibody, which could reverse the inhibited lymphocyte proliferation by the immuno-suppressive protein. And the antibody could be used to coat plate for ELISA assay. Therefore, the antibody will have the potential to provide a basis for applying to clinical stress diseases in the future.
基金supported by the National Basic Research Development Program(973)of China(No.2009CB5265102010CB529801)+5 种基金National High Technology Research and Development Program(863)of China(No.2006AA02Z141)Natural Science Foundation of Hunan ProvinceChina(No.07JJ3072)Program for New Century Excellent Talents in UniversityMinistry of EducationChina(NCET-07-0279)
