Research Service, Department of Veterans Affairs, Edward Hines Jr. VA Hospital, Hines, IL, USA.
Department of Ophthalmology, Stritch School of Medicine, Loyola University Chicago, Maywood, IL, USA.
Methods Mol Biol. 2024;2816:101-115. doi: 10.1007/978-1-0716-3902-3_10.
Members of the Rho family of small monomeric GTPases regulate a plethora of critical cellular functions including gene expression, cell cycle progression, and the dynamic modeling of the actin cytoskeleton. Diversity among Rho family members is derived, in part, from variations in their subcellular distribution. Localization of newly synthesized (naïve) Rho proteins to target subcellular compartments is largely governed by lipid modifications, including posttranslational prenylation. Here, using well-established and widely available contemporary methodologies, detailed protocols by which to semiquantitatively evaluate the functional consequence of posttranslational prenylation in human trabecular meshwork cells are described. We propose the novel concept that posttranslational prenylation itself is a key regulator of mammalian Rho GTPase protein expression and turnover.
Rho 家族的小单体 GTP 酶成员调节着许多关键的细胞功能，包括基因表达、细胞周期进程和肌动蛋白细胞骨架的动态建模。Rho 家族成员的多样性部分来源于它们在亚细胞分布上的差异。新合成的（幼稚的）Rho 蛋白向靶亚细胞区室的定位在很大程度上受脂质修饰的控制，包括翻译后 prenylation。在这里，使用成熟且广泛可用的当代方法学，我们描述了详细的方案，用于半定量评估人眼小梁组织细胞中转译后 prenylation 的功能后果。我们提出了一个新的概念，即翻译后 prenylation 本身是哺乳动物 Rho GTPase 蛋白表达和周转的关键调节剂。
