Activation of MEKK1 by Rho GTPases

Zhui Chen, Melanie H. Cobb

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Mammalian MAP/ERK kinase kinase 1 (MEKK1) is MAP kinase kinase kinase (MAP3K) that is a crucial regulator of many cellular signaling cascades. One of the most important physiological functions of MEKK1 is its ability to regulate cell migration, because MEKK1 null mice are defective in eyelid closure. MEKK1 exhibits its signaling activity through interaction with a large array of cellular factors, including several proteins that are known to play central roles in controlling cell movement and motility. We have recently identified an interaction between MEKK1 and RhoA. This interaction occurs between the GTP-bound, active form of RhoA and the amino terminal region of MEKK1 that harbors a PHD domain with E3 ubiquitin ligase activity. RhoA-GTP activates MEKK1 in vitro and in cells. Here we describe in detail the assay methods for RhoA activation of MEKK1, including preparation of recombinant proteins and proteins immunoprecipitated from cells, pretreatment of proteins, and assay conditions. We also briefly explain the methods and conditions we use to identify the interaction between MEKK1 and RhoA in yeast and in mammalian cells.

Original languageEnglish (US)
Article number35
Pages (from-to)468-478
Number of pages11
JournalMethods in Enzymology
Volume406
DOIs
StatePublished - 2006

Fingerprint

MAP Kinase Kinase Kinase 1
rho GTP-Binding Proteins
Chemical activation
Cell Movement
Guanosine Triphosphate
Assays
Cells
Cell signaling
MAP Kinase Kinase Kinases
Proteins
Ubiquitin-Protein Ligases
Eyelids
Ports and harbors
Recombinant Proteins
Yeast
Yeasts

ASJC Scopus subject areas

  • Biochemistry

Cite this

Activation of MEKK1 by Rho GTPases. / Chen, Zhui; Cobb, Melanie H.

In: Methods in Enzymology, Vol. 406, 35, 2006, p. 468-478.

Research output: Contribution to journalArticle

Chen, Zhui ; Cobb, Melanie H. / Activation of MEKK1 by Rho GTPases. In: Methods in Enzymology. 2006 ; Vol. 406. pp. 468-478.
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