Analysis of activated GAPs and GEFs in cell lysates

Rafael García-Mata; Krister Wennerberg; William T Arthur; Nicole K Noren; Shawn M Ellerbroek; Keith Burridge

doi:10.1016/S0076-6879(06)06031-9

Analysis of activated GAPs and GEFs in cell lysates

Rafael García-Mata, Krister Wennerberg, William T Arthur, Nicole K Noren, Shawn M Ellerbroek, Keith Burridge

146 Citations (Scopus)

Abstract

An assay was developed that allows the precipitation of the active pools of Rho-GEFs, Rho-GAPs, or effectors from cell or tissue lysates. This assay can be used to identify GEFs, GAPs, and effectors involved in specific cellular pathways to determine their GTPase specificity and to monitor the temporal activation of GEFs and GAPs in response to upstream signals.

Original language	English
Book series	Methods in Enzymology
Volume	406
Pages (from-to)	425-37
Number of pages	13
ISSN	0076-6879
DOIs	https://doi.org/10.1016/S0076-6879(06)06031-9
Publication status	Published - 2006
Externally published	Yes

Keywords

Animals
CHO Cells
Cricetinae
GTPase-Activating Proteins/metabolism
Glutathione Transferase/genetics
Guanine Nucleotide Exchange Factors/metabolism
Humans
Mice
NIH 3T3 Cells
Recombinant Fusion Proteins/metabolism
rho GTP-Binding Proteins/genetics

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10.1016/S0076-6879(06)06031-9

Cite this

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title = "Analysis of activated GAPs and GEFs in cell lysates",

abstract = "An assay was developed that allows the precipitation of the active pools of Rho-GEFs, Rho-GAPs, or effectors from cell or tissue lysates. This assay can be used to identify GEFs, GAPs, and effectors involved in specific cellular pathways to determine their GTPase specificity and to monitor the temporal activation of GEFs and GAPs in response to upstream signals.",

keywords = "Animals, CHO Cells, Cricetinae, GTPase-Activating Proteins/metabolism, Glutathione Transferase/genetics, Guanine Nucleotide Exchange Factors/metabolism, Humans, Mice, NIH 3T3 Cells, Recombinant Fusion Proteins/metabolism, rho GTP-Binding Proteins/genetics",

author = "Rafael Garc{\'i}a-Mata and Krister Wennerberg and Arthur, {William T} and Noren, {Nicole K} and Ellerbroek, {Shawn M} and Keith Burridge",

year = "2006",

doi = "10.1016/S0076-6879(06)06031-9",

language = "English",

volume = "406",

pages = "425--37",

journal = "Methods in Enzymology",

issn = "0076-6879",

publisher = "Academic Press",

}

TY - JOUR

T1 - Analysis of activated GAPs and GEFs in cell lysates

AU - García-Mata, Rafael

AU - Wennerberg, Krister

AU - Arthur, William T

AU - Noren, Nicole K

AU - Ellerbroek, Shawn M

AU - Burridge, Keith

PY - 2006

Y1 - 2006

N2 - An assay was developed that allows the precipitation of the active pools of Rho-GEFs, Rho-GAPs, or effectors from cell or tissue lysates. This assay can be used to identify GEFs, GAPs, and effectors involved in specific cellular pathways to determine their GTPase specificity and to monitor the temporal activation of GEFs and GAPs in response to upstream signals.

AB - An assay was developed that allows the precipitation of the active pools of Rho-GEFs, Rho-GAPs, or effectors from cell or tissue lysates. This assay can be used to identify GEFs, GAPs, and effectors involved in specific cellular pathways to determine their GTPase specificity and to monitor the temporal activation of GEFs and GAPs in response to upstream signals.

KW - Animals

KW - CHO Cells

KW - Cricetinae

KW - GTPase-Activating Proteins/metabolism

KW - Glutathione Transferase/genetics

KW - Guanine Nucleotide Exchange Factors/metabolism

KW - Humans

KW - Mice

KW - NIH 3T3 Cells

KW - Recombinant Fusion Proteins/metabolism

KW - rho GTP-Binding Proteins/genetics

U2 - 10.1016/S0076-6879(06)06031-9

DO - 10.1016/S0076-6879(06)06031-9

M3 - Journal article

C2 - 16472675

SN - 0076-6879

VL - 406

SP - 425

EP - 437

JO - Methods in Enzymology

JF - Methods in Enzymology

ER -

Analysis of activated GAPs and GEFs in cell lysates

Abstract

Keywords

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