Croft Daniel R, Olson Michael F
The Beatson Institute for Cancer Research, Glasgow, UK.
Methods Enzymol. 2006;406:541-53. doi: 10.1016/S0076-6879(06)06042-3.
The Rho-associated kinases ROCK I and ROCK II are serine/threonine kinases that play central and critical roles in regulating the actin cytoskeleton. Activation of ROCK proteins contributes positively to the phosphorylation of myosin II light chains (MLC), myosin ATPase activity, stabilization of filamentous actin, and coupling of the actin-myosin filaments to the plasma membrane, thereby leading to the increased actin-myosin force generation and contractility. We have constructed a conditionally-activated form of ROCK II (called ROCK:ER) by fusing the ROCK II kinase domain to the estrogen receptor hormone-binding domain. In this chapter, we describe the construction and characterization of this regulatable ROCK:ER fusion protein.
Rho相关激酶ROCK I和ROCK II是丝氨酸/苏氨酸激酶，在调节肌动蛋白细胞骨架中发挥核心和关键作用。ROCK蛋白的激活对肌球蛋白轻链（MLC）的磷酸化、肌球蛋白ATP酶活性、丝状肌动蛋白的稳定以及肌动蛋白-肌球蛋白丝与质膜的偶联有积极作用，从而导致肌动蛋白-肌球蛋白产生的力和收缩性增加。我们通过将ROCK II激酶结构域与雌激素受体激素结合结构域融合，构建了一种条件激活形式的ROCK II（称为ROCK:ER）。在本章中，我们描述了这种可调节的ROCK:ER融合蛋白的构建和特性。
