Stubbs Evan B
Research Service, Department of Veterans Affairs, Edward Hines Jr. VA Hospital, Hines, IL, 60141, USA.
Department of Ophthalmology, Stritch School of Medicine, Loyola University Chicago, 2160 South First Ave., LUH-North Entrance, Suite 2601, Maywood, IL, 60153, USA.
Methods Mol Biol. 2017;1609:217-229. doi: 10.1007/978-1-4939-6996-8_18.
Small monomeric GTPases, including those belonging to the Rho family, regulate a diverse array of intracellular signaling pathways which affect vesicle transport/trafficking, endocytosis, cell cycle progression, cell contractility, and formation of stress fibers or focal adhesions. Functional activation of newly synthesized small monomeric GTPases is facilitated by a multistep post-translational process involving transferase-catalyzed addition of farnesyl or geranylgeranyl isoprenoids to conserved cysteine residues within a unique carboxy terminal CaaX motif. Here, using well-established and widely available contemporary methodologies, detailed protocols by which to semi-quantitatively evaluate the functional consequence of post-translational isoprenylation in human trabecular meshwork cells are described. We introduce the concept that isoprenylation alone is itself a key regulator of mammalian Rho GTPase expression and turnover.
小的单体GTP酶，包括那些属于Rho家族的成员，调节着一系列不同的细胞内信号通路，这些通路影响囊泡运输/ trafficking、内吞作用、细胞周期进程、细胞收缩性以及应力纤维或粘着斑的形成。新合成的小单体GTP酶的功能激活通过一个多步骤的翻译后过程来促进，该过程涉及转移酶催化的将法尼基或香叶基香叶基类异戊二烯添加到独特的羧基末端CaaX基序内保守的半胱氨酸残基上。在这里，使用成熟且广泛可用的当代方法，描述了用于半定量评估人小梁网细胞中翻译后异戊二烯化功能后果的详细方案。我们提出了这样一个概念，即仅异戊二烯化本身就是哺乳动物Rho GTP酶表达和周转的关键调节因子。
