Department of Biochemistry, Stanford University School of Medicine, Stanford, CA, USA.
Methods Mol Biol. 2021;2293:19-25. doi: 10.1007/978-1-0716-1346-7_2.
Rab29 has been implicated in multiple membrane trafficking processes with no described effectors or regulating proteins. Its fast nucleotide exchange rate and inability to bind GDI in cytosol make it a unique and poorly understood Rab. Because the conventional, "GTP-locked" Rab mutation does not have the desired effect in Rab29, we present here the use of a fluorescence-based assay to characterize novel Rab29 mutants (I64T and V156G) that display faster nucleotide exchange rates, allowing for GEF-independent Rab29 activation.
Rab29 参与了多种膜运输过程,但目前尚不清楚其效应物或调节蛋白。由于常规的“GTP 锁定”Rab 突变在 Rab29 中没有达到预期效果,因此我们在此提出使用基于荧光的测定法来表征具有更快核苷酸交换率的新型 Rab29 突变体(I64T 和 V156G),从而实现 GEF 独立的 Rab29 激活。Rab29 的快速核苷酸交换率和在细胞质中不能结合 GDI 使其成为一种独特且了解甚少的 Rab。