Methods to Study the Unique SOCS Box Domain of the Rab40 Small GTPase Subfamily.
Methods to Study the Unique SOCS Box Domain of the Rab40 Small GTPase Subfamily.
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DOI：
10.1007/978-1-0716-1346-7_11
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Prekeris R
中科院分区：
文献类型：
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作者：
Duncan ED;Lencer E;Linklater E;Prekeris R
尽管Rab GTP酶在细胞内转运中起着关键作用，但该家族中的绝大多数蛋白质仍缺乏特征性，包括Rab 40亚家族。Rab 40蛋白家族通常被认为是非典型Rab，与任何其他小GTPase不同，因为它们包含细胞因子信号传导（SOCS）盒的C末端抑制因子。已经确定，其他蛋白质中的该SOCS结构域介导与支架蛋白Cullin 5的相互作用，以形成对蛋白质泛素化和周转至关重要的E3泛素连接酶复合物。虽然SOCS/Cullin 5复合物的功能已经在这些其他蛋白质中的几个中得到了很好的定义，但Rab 40蛋白质家族的情况还不是这样。我们之前已经证明Rab 40 b家族成员在三维（3D）乳腺癌细胞迁移过程中起着重要作用。为了进一步了解这一点，我们开始研究Rab 40 b在细胞迁移过程中的SOCS依赖性作用。在这里，我们描述了一个公正的方法来确定潜在的Rab 40 b/Cullin 5底物。我们预计，这种方法将是有用的，研究其他Rab 40家族成员以及其他SOCS盒蛋白的功能。
Despite the critical role of Rab GTPases for intracellular transport, the vast majority of proteins within this family remain poorly characterized, including the Rab40 subfamily. Often recognized as atypical Rabs, the Rab40 family of proteins are unlike any other small GTPase because they contain a C-terminal suppressor of cytokine signaling (SOCS) box. It is well established that this SOCS domain in other proteins mediates an interaction with the scaffold protein Cullin5 in order to form a E3 ubiquitin ligase complex critical for protein ubiquitylation and turnover. Although the function of SOCS/Cullin5 complexes has been well defined in several of these other proteins, this is not yet the case for the Rab40 family of proteins. We have previously shown that the Rab40b family member plays an important role during three-dimensional (3D) breast cancer cell migration. To further this knowledge, we began to investigate the SOCS-dependent role of Rab40b during cell migration. Here, we describe an unbiased approach to identify potential Rab40b/Cullin5 substrates. We anticipate that this method will be useful for studying the function of other Rab40 family members as well as other SOCS box containing proteins.