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Regulators and effectors of small GTPases [electronic resource] : Rho family / edited by William E. Balch, Channing J. Der, Alan Hall.

Contributor(s): Balch, William Edward, 1949- | Der, Channing J | Hall, A. (Alan)Material type: TextTextSeries: Methods in enzymology ; v. 406.Publication details: Amsterdam ; Boston : Elsevier : Academic Press, c2006. Description: 1 online resource (xliii, 798 p.) : illISBN: 0080479391 (electronic bk.); 9780080479392 (electronic bk.); 0121828115 (electronic bk.); 9780121828110 (electronic bk.)Subject(s): Guanosine triphosphatase | Rho GTPases | rho GTP-Binding Proteins -- physiology | GTPasen | Regelmechanismen | SCIENCE / Life Sciences / Molecular Biology | Guanosine triphosphatase | Rho GTPasesGenre/Form: Electronic books.DDC classification: 572.8 LOC classification: QP601 | .C57 v. 406NLM classification: W1 | QU 55Other classification: 35.74 | 35.70 Online resources: ScienceDirect | ScienceDirect | Volltext
Contents:
Purification and Biochemical Properties of Rac1, 2, 3 and the Splice Variant Rac1b -- Biochemical Analyses of the Wrch Atypical Rho Family GTPases -- Purification of P-Rex1 from Neutrophils and Nucleotide Exchange Assay -- In Vitro Guanine Nucleotide Exchange Activity of DHR-2/DOCKER/CZH2 Domains -- Biochemical Characterization of the Cool (Cloned-Out-of-Library)/Pix (Pak-Interactive Exchange Factor) Proteins -- GEF and Glucosylation Assays on Liposome-Bound Rac -- Phosphorylation of RhoGDI by p21-activated kinase 1 -- Purification of ARAP3 and Characterization of GAP Activities -- Regulation of RhoGAP specificity by Phospholipids and Prenylation -- Purification and Activity of the Rho ADP-ribosylating binary C2/C3 Toxin -- Purification of TAT-C3 Exoenzyme -- Imaging and photobleach correction of Mero-CBD, Sensor of Endogenous Cdc42 Activation -- Cdc42 and PI(4,5)P2-induced actin Assembly in Xenopus Egg Extracts -- In vitro reconstitution of Cdc42-mediated actin assembly using purified components -- Biochemical analysis of mammalian formin effects on actin Dynamics -- Formin proteins: purification and measurement of effects on actin assembly -- Purification and kinase assay of PKN -- Purification and enzyme activity of ACK1 -- Direct Activation of purified phospholipase C epsilon by RhoA studied in reconstituted phospholipid Vesicles -- Regulation of PLCbeta isoforms by Rac -- Biochemical properties and Inhibitors of (N- )WASP -- The Use of GFP to localize Rho GTPases in living cells -- Analysis of the spatiotemporal activation of Rho GTPases using Raichu probes -- Measurement of activity of Rho GTPases during mitosis -- Inhibition of Rho GTPases by RNA interference -- RNA interference techniques to study epithelial cell adhesion and polarity -- Nucleofection of Primary Neurons -- Dock180-ELMO cooperation in Rac activation -- Rho GTPase activation by cell-cell adhesion -- Activation of Rap1, Cdc42, and Rac by nectin adhesion system -- Analysis of activated GAPs and GEFs in cell lysates -- Degradation of RhoA by Smurf1 ubiquitin ligase -- Ubiquitin-mediated proteasomal degradation of Rho proteins by the CNF1 toxin -- Regulation of superoxide-producing NADPH oxidases in nonphagocytic cells -- Activation of MEKK1 by Rho GTPases -- Activation of the apoptotic JNK pathway through the Rac1-binding scaffold protein POSH -- Quantification of isozyme-specific activation of phospholipase C-beta2 by Rac GTPases and phospholipase C-epsiv by Rho GTPases in an intact cell assay system -- Activiation of Rho and Rac by Wnt/frizzled signaling -- Fluorescent assay of cell-permeable C3 transfease activity -- Use of TIRF microscopy to visualize actin and microtubules in migrating cells -- Inhibition of ROCK by RhoE -- Conditional Regulation of a ROCK-estrogen receptor fusion protein -- Rational design and applications of a Rac GTPase-specific small molecule Inhibitor -- In vitro assay of primary astrocyte migration as a tool to study Rho GTPase function in cell polarization -- Real-time centrosome reorientation during fibroblast migration -- Lentiviral delivery of RNAi in hippocampal neurons -- Methods for studying neutrophil chemotaxis -- Measurement of epidermal growth factor receptor turnover and effects of Cdc42 -- Tumor cell migration in three dimensions -- An in vitro model to study the role of endothelial Rho GTPases during leukocyte transendothelial migration -- Analysis of a mitotic role of Cdc42 -- Plexin-induced collapse assay in COS cells -- Morphological and biochemical analysis of Rac1 in three-dimensional epithelial cell cultures -- Using three-dimensional acinar structures for molecular and cell biological assays -- TC10 and insulin-stimulated glucose transport -- GTPases and the control of neuronal polarity -- In vitro assembly of filopodia-like bundles.
Summary: The Ras superfamily (>150 human members) encompasses Ras GTPases involved in cell proliferation, Rho GTPases involved in regulating the cytoskeleton, Rab GTPases involved in membrane targeting/fusion and a group of GTPases including Sar1, Arf, Arl and dynamin involved in vesicle budding/fission. These GTPases act as molecular switches and their activities are controlled by a large number of regulatory molecules that affect either GTP loading (guanine nucleotide exchange factors or GEFs) or GTP hydrolysis (GTPase activating proteins or GAPs). In their active state, they interact with a continua.
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Includes bibliographical references and indexes.

Purification and Biochemical Properties of Rac1, 2, 3 and the Splice Variant Rac1b -- Biochemical Analyses of the Wrch Atypical Rho Family GTPases -- Purification of P-Rex1 from Neutrophils and Nucleotide Exchange Assay -- In Vitro Guanine Nucleotide Exchange Activity of DHR-2/DOCKER/CZH2 Domains -- Biochemical Characterization of the Cool (Cloned-Out-of-Library)/Pix (Pak-Interactive Exchange Factor) Proteins -- GEF and Glucosylation Assays on Liposome-Bound Rac -- Phosphorylation of RhoGDI by p21-activated kinase 1 -- Purification of ARAP3 and Characterization of GAP Activities -- Regulation of RhoGAP specificity by Phospholipids and Prenylation -- Purification and Activity of the Rho ADP-ribosylating binary C2/C3 Toxin -- Purification of TAT-C3 Exoenzyme -- Imaging and photobleach correction of Mero-CBD, Sensor of Endogenous Cdc42 Activation -- Cdc42 and PI(4,5)P2-induced actin Assembly in Xenopus Egg Extracts -- In vitro reconstitution of Cdc42-mediated actin assembly using purified components -- Biochemical analysis of mammalian formin effects on actin Dynamics -- Formin proteins: purification and measurement of effects on actin assembly -- Purification and kinase assay of PKN -- Purification and enzyme activity of ACK1 -- Direct Activation of purified phospholipase C epsilon by RhoA studied in reconstituted phospholipid Vesicles -- Regulation of PLCbeta isoforms by Rac -- Biochemical properties and Inhibitors of (N- )WASP -- The Use of GFP to localize Rho GTPases in living cells -- Analysis of the spatiotemporal activation of Rho GTPases using Raichu probes -- Measurement of activity of Rho GTPases during mitosis -- Inhibition of Rho GTPases by RNA interference -- RNA interference techniques to study epithelial cell adhesion and polarity -- Nucleofection of Primary Neurons -- Dock180-ELMO cooperation in Rac activation -- Rho GTPase activation by cell-cell adhesion -- Activation of Rap1, Cdc42, and Rac by nectin adhesion system -- Analysis of activated GAPs and GEFs in cell lysates -- Degradation of RhoA by Smurf1 ubiquitin ligase -- Ubiquitin-mediated proteasomal degradation of Rho proteins by the CNF1 toxin -- Regulation of superoxide-producing NADPH oxidases in nonphagocytic cells -- Activation of MEKK1 by Rho GTPases -- Activation of the apoptotic JNK pathway through the Rac1-binding scaffold protein POSH -- Quantification of isozyme-specific activation of phospholipase C-beta2 by Rac GTPases and phospholipase C-epsiv by Rho GTPases in an intact cell assay system -- Activiation of Rho and Rac by Wnt/frizzled signaling -- Fluorescent assay of cell-permeable C3 transfease activity -- Use of TIRF microscopy to visualize actin and microtubules in migrating cells -- Inhibition of ROCK by RhoE -- Conditional Regulation of a ROCK-estrogen receptor fusion protein -- Rational design and applications of a Rac GTPase-specific small molecule Inhibitor -- In vitro assay of primary astrocyte migration as a tool to study Rho GTPase function in cell polarization -- Real-time centrosome reorientation during fibroblast migration -- Lentiviral delivery of RNAi in hippocampal neurons -- Methods for studying neutrophil chemotaxis -- Measurement of epidermal growth factor receptor turnover and effects of Cdc42 -- Tumor cell migration in three dimensions -- An in vitro model to study the role of endothelial Rho GTPases during leukocyte transendothelial migration -- Analysis of a mitotic role of Cdc42 -- Plexin-induced collapse assay in COS cells -- Morphological and biochemical analysis of Rac1 in three-dimensional epithelial cell cultures -- Using three-dimensional acinar structures for molecular and cell biological assays -- TC10 and insulin-stimulated glucose transport -- GTPases and the control of neuronal polarity -- In vitro assembly of filopodia-like bundles.

The Ras superfamily (>150 human members) encompasses Ras GTPases involved in cell proliferation, Rho GTPases involved in regulating the cytoskeleton, Rab GTPases involved in membrane targeting/fusion and a group of GTPases including Sar1, Arf, Arl and dynamin involved in vesicle budding/fission. These GTPases act as molecular switches and their activities are controlled by a large number of regulatory molecules that affect either GTP loading (guanine nucleotide exchange factors or GEFs) or GTP hydrolysis (GTPase activating proteins or GAPs). In their active state, they interact with a continua.

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