Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, 3333 Burnet Avenue, Cincinnati, OH 45229, USA.
NMR-based Metabolomics Core, Division of Pathology and Laboratory Medicine, Cincinnati Children's Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, OH 45229, USA.
STAR Protoc. 2021 May 14;2(2):100541. doi: 10.1016/j.xpro.2021.100541. eCollection 2021 Jun 18.
Ras GTPases in complex with Guanosine triphosphate (GTP) or GTP analog exhibit dynamic equilibrium between two interconvertible conformations-an inactive state 1 and an active state 2. Unlike Ras, it remains unclear if the GTP-bound form of Rho GTPases also exhibits multiple conformational states. Here, we describe a protocol for structural and biochemical analyses of RhoA GTPase. This protocol can be adapted for the characterization of other Rho GTPases. For details on the use and execution of this protocol, please refer to Lin et al. (2021).
Ras GTPases 与鸟苷三磷酸(GTP)或 GTP 类似物形成复合物后,在两种可相互转化的构象之间表现出动态平衡——一种是无活性状态 1,另一种是活性状态 2。与 Ras 不同,目前尚不清楚 Rho GTPases 的 GTP 结合形式是否也表现出多种构象状态。在这里,我们描述了一种用于 RhoA GTPase 结构和生化分析的方案。该方案可适用于其他 Rho GTPases 的特征描述。有关此方案的使用和执行的详细信息,请参见 Lin 等人。(2021)。