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Regulators and Effectors of Small GTPases: Rho Family
W. E. Balch (Volume editor), Channing J. Der (Volume editor), Alan Hall (Volume editor)
9780121828110, Elsevier Science
Hardback, published 21 February 2006
848 pages, Approx. 250 illustrations (50 in full color)
22.9 x 15.1 x 4.2 cm, 1.29 kg
Praise for the Series "Incomparably useful." --ANALYTICAL BIOCHEMISTRY "It is a true 'methods' series, including almost every detail from basic theory to sources of equipment and reagents, with timely documentation provided on each page." --BIO/TECHNOLOGY "The series has been following the growing, changing and creation of new areas of science. It should be on the shelves of all libraries in the world as a whole collection." --CHEMISTRY IN INDUSTRY
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 continually increasing, functionally complex array of downstream effectors. Since the last Methods in Enzymology volume on this topic in 2000, Rho GTPases have continued to receive a huge amount of attention. The human genome sequence has revealed the full extent of the Rho GEF and Rho GAP families (over 80 members for each) and the challenge of identifying the molecular interactions and cellular pathways influenced by each of these regulators is a daunting prospect. This new volume, Regulators and Effectors of Small GTPases: Rho Family, describes some of the methods currently being used to examine Rho family GTPase regulation at the biochemical and cellular level.
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 (Coned-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 exoenzymes
Imaging and photobleach correction of MeroCBD, 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 PLCâ isoforms by Rac
Biochemical properties and inhibitors of (N-)WASP
The Use of GFP to localize Rho GTPases in Living Cells
Analysis of the Spatio-Temporal Activation of Rho GTPases using Raichu Probes
Measurement of Activity of Rho GTPases during Mitosis
RNAi inhibition of Rho GTPases
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 in non-phagocytic cells
Activation of MEKK1 by Rho GTPases
Activation of Apoptotic JNK Pathway through the Rac1-Binding Scaffold Protein POSH
Quantification of isozyme-specific activation of phospholipase C-â2 by Rac GTPases and phospholipase C-å by Rho GTPases in an intact cell assay system
Activation of Rho and Rac by Wnt/Frizzled signalling
Fluorescent assay of cell permeable C3 transferase 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 Chemtoaxis
Measurement of Epidermal Growth Factor Receptor turnover and effects of Cdc42
Tumor cell migration in three dimensions
Reciprocal regulation of cyclin D1 expression by Rac and Rho
Regulation of ERK signal duration and cyclin D1 expression by Rhodependent stress fiber formation
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
Subject Areas: Cellular biology [cytology PSF], Molecular biology [PSD], Proteins [PSBC], Biochemistry [PSB], Biophysics [PHVN]
