GTPases Regulating Membrane Targeting and Fusion

GTPases Regulating Membrane Targeting and Fusion

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Rab GTPases now comprise a family of g63 members. They are emerging as the key hub element controlling the membrane architecture of eukaryotic cells. They are intimately involved in vesicle targeting and fusion in both the endocytic and exocytic pathways and direct the assembly and disassembly of protein complexes that include regulators (GEFs and GAPs), effectors (tethers/motors) and fusion components (SNAREs) that control membrane targeting and fusion. During the last 3 years the field has virtually exploded with the identification and characterization of many new Rab proteins and their effectors. Our understanding of how Rab GTPases control membrane function remains at its infancy. This volume of MIE provides a wealth of new concepts, approaches and tools to study Rab proteins in the test tube and in living cells that will be of strong benefit to both established laboratories and new investigators in the field to elucidate Rab GTPase function in cellular development, differentiation and proliferation. * Comprehensive overview of Rab GTPase phylogeny and systems biology * Identification and characterization of Rab GEFs, GAPs and effectors * General methodologies to study Rab GTPase function in vitro and in vivo using biochemical, molecular and microscopy approachespH 7.5, 200 mM NaCl, 20 mM EDTA, 0.1% [v/v] Triton X-100) rotating at 4Ad for 5- 10 min. Pellet the beads by centrifu- gation at 2000 xg for 1 min and then transfer the supernatant to a fresh tube, avoiding the beads. An optional step at this ... for 5 min, 4Ad. The acetone washing steps help to remove the residual TCA, which otherwise causes problems for the solubi- lization of the precipitated protein. Removeanbsp;...

Title:GTPases Regulating Membrane Targeting and Fusion
Author:William Edward Balch, Channing J. Der, Alan Hall
Publisher:Gulf Professional Publishing - 2005


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