棒状体相关蛋白1(rhoptry-associated protein 1,RAP-1)是由棒状体分泌的与侵袭宿主细胞相关的蛋白。本研究以顶复门原虫RAP-1基因为靶标,用最大似然法构建了巴贝斯属、疟原虫属、泰勒虫属的RAP-1基因进化树;应用软件预测分析RAP-1蛋白...棒状体相关蛋白1(rhoptry-associated protein 1,RAP-1)是由棒状体分泌的与侵袭宿主细胞相关的蛋白。本研究以顶复门原虫RAP-1基因为靶标,用最大似然法构建了巴贝斯属、疟原虫属、泰勒虫属的RAP-1基因进化树;应用软件预测分析RAP-1蛋白的理化性质、亲疏水性、跨膜区以及二、三级结构,并表达纯化了马泰勒虫(Theileria equi)RAP-1重组蛋白。结果显示:巴贝斯属不同虫株相聚类,测定序列与马泰勒虫聚为一支,又与恶性疟原虫相聚类,表明亲缘关系较近。马泰勒虫RAP-1蛋白理论等电点为9.85,半衰期为30 h,总平均亲水性(GRAVY)为-0.368,无跨膜区;RAP-1蛋白二级结构中,α-螺旋、β-折叠、β-转角和无规卷曲占比分别为38.5%、15.5%、27.8%、18.1%;构建的三级结构立体展现了RAP-1蛋白形态。成功构建了原核表达载体pET-32a-RAP-1;该重组蛋白主要以包涵体形式存在,蛋白大小为65 kDa。RAP-1蛋白可通过原核表达系统高效表达,纯化后的产物能被马泰勒虫标准阳性血清识别,具有良好的反应原性。本研究为深入了解马泰勒虫入侵宿主的机制机理以及RAP-1蛋白的功能研究奠定了基础。展开更多
Rap1A is a small G protein implicated in a spectrum of biological processes such as cell proliferation,adhesion,differentiation,and embryogenesis.The downstream effectors through which Rap1A mediates its diverse effec...Rap1A is a small G protein implicated in a spectrum of biological processes such as cell proliferation,adhesion,differentiation,and embryogenesis.The downstream effectors through which Rap1A mediates its diverse effects are largely unknown.Here we show that Rap1A,but not the related small G proteins Rap2 or Ras,binds the tumor suppressor Ras association domain family 1A(RASSF1A)in a manner that is regulated by phosphorylation of RASSF1A.Interaction with Rap1A is shown to influence the effect of RASSF1A on microtubule behavior.展开更多
基金SKV also acknowledges the award of the Commonwealth Scholarship and the financial support received from the Department of Medical Oncology,Medical Sciences Division,The University of Oxford,Oxford,UK and the Cancer Research UK.
文摘Rap1A is a small G protein implicated in a spectrum of biological processes such as cell proliferation,adhesion,differentiation,and embryogenesis.The downstream effectors through which Rap1A mediates its diverse effects are largely unknown.Here we show that Rap1A,but not the related small G proteins Rap2 or Ras,binds the tumor suppressor Ras association domain family 1A(RASSF1A)in a manner that is regulated by phosphorylation of RASSF1A.Interaction with Rap1A is shown to influence the effect of RASSF1A on microtubule behavior.
