Rho GTPases: Secretion and actin dynamics in permeabilized mast cells
This chapter describes the Rho guanosine 5'-triphosphates (GTPases) secretion and actin dynamics in permeabilized mast cells. Permeabilized mast cells proved to be a valuable tool for studying the function of Rho-related GTPases. Rho, Rac, and Cdc42 were shown to be crucial for regulation of th...
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| Veröffentlicht in: | Methods in Enzymology 2000, Vol.325, p.356-369 |
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| creator | Holt, Mark R. Koffer, Anna |
| description | This chapter describes the Rho guanosine 5'-triphosphates (GTPases) secretion and actin dynamics in permeabilized mast cells. Permeabilized mast cells proved to be a valuable tool for studying the function of Rho-related GTPases. Rho, Rac, and Cdc42 were shown to be crucial for regulation of the late steps of exocytosis. The chapter discusses the use of primary mast cells—permeabilized by streptolysin O (SLO)—for studying Rho family GTPase. The chapter discusses the preparation of recombinant GTPases, an assay for GTP-binding activity, and subsequent loading of recombinant proteins with GTP. The chapter describes various procedures for introducing GTPase into mast cells and evaluation of their effects on the cytoskeleton and exocytosis. The chapter also discusses the isolation, permeabilization, and stimulation of rat peritoneal mast cells. The chapter concludes with a discussion of the analysis of F-actin content and morphology. |
| doi_str_mv | 10.1016/S0076-6879(00)25457-8 |
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| ispartof | Methods in Enzymology, 2000, Vol.325, p.356-369 |
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| subjects | Actins - metabolism ADP Ribose Transferases - genetics ADP Ribose Transferases - metabolism Animals Bacterial Proteins Botulinum Toxins Cell Membrane - drug effects Cell Membrane Permeability Exocytosis Guanosine Triphosphate - metabolism Mast Cells - metabolism Peritoneal Cavity - cytology Rats Recombinant Fusion Proteins - metabolism rho GTP-Binding Proteins - genetics rho GTP-Binding Proteins - metabolism Streptolysins - pharmacology |
| title | Rho GTPases: Secretion and actin dynamics in permeabilized mast cells |
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