Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Reconstitution Of β-catenin degradation in Xenopus egg extract.
Chen TW
,
Broadus MR
,
Huppert SS
,
Lee E
.
???displayArticle.abstract???
Xenopus laevis egg extract is a well-characterized, robust system for studying the biochemistry of diverse cellular processes. Xenopus egg extract has been used to study protein turnover in many cellular contexts, including the cell cycle and signal transduction pathways(1-3). Herein, a method is described for isolating Xenopus egg extract that has been optimized to promote the degradation of the critical Wnt pathway component, β-catenin. Two different methods are described to assess β-catenin protein degradation in Xenopus egg extract. One method is visually informative ([(35)S]-radiolabeled proteins), while the other is more readily scaled for high-throughput assays (firefly luciferase-tagged fusion proteins). The techniques described can be used to, but are not limited to, assess β-catenin protein turnover and identify molecular components contributing to its turnover. Additionally, the ability to purify large volumes of homogenous Xenopus egg extract combined with the quantitative and facile readout of luciferase-tagged proteins allows this system to be easily adapted for high-throughput screening for modulators of β-catenin degradation.
???displayArticle.pubmedLink???
24962160
???displayArticle.pmcLink???PMC4133086 ???displayArticle.link???J Vis Exp ???displayArticle.grants???[+]
Blow,
Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs.
1986, Pubmed,
Xenbase
Blow,
Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs.
1986,
Pubmed
,
Xenbase
Chan,
In vitro study of nuclear assembly and nuclear import using Xenopus egg extracts.
2006,
Pubmed
,
Xenbase
Cross,
Preparation and fractionation of Xenopus laevis egg extracts.
2008,
Pubmed
,
Xenbase
Cross,
Obtaining eggs from Xenopus laevis females.
2008,
Pubmed
,
Xenbase
Cselenyi,
LRP6 transduces a canonical Wnt signal independently of Axin degradation by inhibiting GSK3's phosphorylation of beta-catenin.
2008,
Pubmed
,
Xenbase
Dabauvalle,
Assembly of nuclear pore complexes in Xenopus egg extract.
1991,
Pubmed
,
Xenbase
Forbes,
Spontaneous formation of nucleus-like structures around bacteriophage DNA microinjected into Xenopus eggs.
1983,
Pubmed
,
Xenbase
Glotzer,
Cyclin is degraded by the ubiquitin pathway.
1991,
Pubmed
,
Xenbase
Guger,
beta-Catenin has Wnt-like activity and mimics the Nieuwkoop signaling center in Xenopus dorsal-ventral patterning.
1995,
Pubmed
,
Xenbase
Hinkson,
The dynamic state of protein turnover: It's about time.
2011,
Pubmed
Jernigan,
Gbetagamma activates GSK3 to promote LRP6-mediated beta-catenin transcriptional activity.
2010,
Pubmed
,
Xenbase
Kinzler,
Lessons from hereditary colorectal cancer.
1996,
Pubmed
Kornbluth,
Analysis of the cell cycle using Xenopus egg extracts.
2006,
Pubmed
,
Xenbase
Lee,
Physiological regulation of [beta]-catenin stability by Tcf3 and CK1epsilon.
2001,
Pubmed
,
Xenbase
Lee,
The roles of APC and Axin derived from experimental and theoretical analysis of the Wnt pathway.
2003,
Pubmed
,
Xenbase
Lohka,
Formation in vitro of sperm pronuclei and mitotic chromosomes induced by amphibian ooplasmic components.
1983,
Pubmed
,
Xenbase
Major,
Wilms tumor suppressor WTX negatively regulates WNT/beta-catenin signaling.
2007,
Pubmed
,
Xenbase
Maresca,
Methods for studying spindle assembly and chromosome condensation in Xenopus egg extracts.
2006,
Pubmed
,
Xenbase
Masui,
Cytoplasmic control of nuclear behavior during meiotic maturation of frog oocytes.
1971,
Pubmed
Mitchison,
Dynamic instability of microtubule growth.
,
Pubmed
Murray,
Cell cycle extracts.
1991,
Pubmed
Newport,
Regulation of the cell cycle during early Xenopus development.
1984,
Pubmed
,
Xenbase
Rubinfeld,
Binding of GSK3beta to the APC-beta-catenin complex and regulation of complex assembly.
1996,
Pubmed
Salic,
Control of beta-catenin stability: reconstitution of the cytoplasmic steps of the wnt pathway in Xenopus egg extracts.
2000,
Pubmed
,
Xenbase
Salic,
Identifying small molecule inhibitors of the ubiquitin-proteasome pathway in Xenopus egg extracts.
2005,
Pubmed
,
Xenbase
Seeling,
Regulation of beta-catenin signaling by the B56 subunit of protein phosphatase 2A.
1999,
Pubmed
,
Xenbase
Shennan,
Xenopus egg extracts: a model system to study proprotein convertases.
2006,
Pubmed
,
Xenbase
Tan,
Wnt signalling pathway parameters for mammalian cells.
2012,
Pubmed
,
Xenbase
Theriot,
Involvement of profilin in the actin-based motility of L. monocytogenes in cells and in cell-free extracts.
1994,
Pubmed
,
Xenbase
Thorne,
Small-molecule inhibition of Wnt signaling through activation of casein kinase 1α.
2010,
Pubmed
,
Xenbase
Thorne,
A biochemical screen for identification of small-molecule regulators of the Wnt pathway using Xenopus egg extracts.
2011,
Pubmed
,
Xenbase
Trinkle-Mulcahy,
Identifying specific protein interaction partners using quantitative mass spectrometry and bead proteomes.
2008,
Pubmed
Tutter,
Chromosomal DNA replication in a soluble cell-free system derived from Xenopus eggs.
2006,
Pubmed
,
Xenbase
Verma,
Ubistatins inhibit proteasome-dependent degradation by binding the ubiquitin chain.
2004,
Pubmed
,
Xenbase
Willis,
Study of the DNA damage checkpoint using Xenopus egg extracts.
2012,
Pubmed
,
Xenbase
Yu,
Identification of a novel ubiquitin-conjugating enzyme involved in mitotic cyclin degradation.
1996,
Pubmed
,
Xenbase
