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Author:
Date:
2020-05-05
[Abstract] The Ras homologous protein (Rho) GTPase subfamily, including RhoA, RhoB, and RhoC are small molecules (~21 kDa) that act as molecular switches in a wide range of signaling pathways to orchestrate biological processes associated with both physiological and tumorigenic cellular states. The Rho GTPases are crucial regulators of actin cytoskeleton rearrangements and FA dynamics and are required for effective cell migration and invasion, as well as cell cycle progression and apoptosis. The Rho GTPases activity is regulated by conformational switching between GTP-bound (active) and GDP-bound (inactive) states. This GTP/GDP cycling is tightly controlled by the guanine nucleotide exchange factors (GEFs), which function as activators by catalyzing the exchange of GDP for GTP and by the ...
[摘要] [摘要] 包括RhoA，RhoB和RhoC在内的Ras同源蛋白（Rho）GTPase亚家族是小分子（〜21 kDa ），在广泛的信号传导途径中充当分子开关，以协调与生理和致瘤细胞状态相关的生物学过程。该Rho GTP酶是至关重要的调节肌动蛋白细胞骨架重排和FA动态和所需要的有效细胞迁移和侵袭，以及细胞周期进程和凋亡，其Rho GTP酶的活动是监管下的构象之间切换GTP结合（有源）和GDP -- GTP / GDP循环受鸟嘌呤核苷酸交换因子（GEF）严格控制，鸟嘌呤核苷酸交换因子（GEF）通过催化GTP的GDP交换和GTPase激活蛋白（GAP）来激活，从而实现水解作用。导致Rho GTPase失活的GTP 的详细信息。在此，我们描述了执行RhoB G-LISA活化测定以检测体外GTP负载的RhoB的水平的详细协议。这是第一个专门用于测量RhoB活化的比色测定。该方法是通过改编RhoA G-LISA激活检测试剂盒（Cytoskeleton，Inc.）而开发的，可在不到3 小时的时间内精确测量RhoB活性，这种快速方法可广泛用于评估GTP负载的RhoB 的水平在任何种类的细胞模型中，要了解RhoB激活在生理过程，疾病，致癌转化中的作用或在高通量筛选中发现药物的作用。
[背景 ] 尽管Rho GTP酶的RhoA，RhoB的，和RhoC占有率超过85 ...
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