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.
Analysis of Transforming Growth Factor ß Family Cleavage Products Secreted Into the Blastocoele of Xenopus laevis Embryos.
Kim HS
,
Christian JL
.
???displayArticle.abstract???
The two arms of the Transforming Growth Factor ß (Tgfß) superfamily, represented by Tgfß/Nodal or Bone morphogenetic protein (Bmp) ligands, respectively, play essential roles in embryonic development and adult homeostasis. Members of the Tgfß family are made as inactive precursors that dimerize and fold within the endoplasmic reticulum. The precursor is subsequently cleaved into ligand and prodomain fragments. Although only the dimeric ligand can engage Tgfß receptors and activate downstream signaling, there is growing recognition that the prodomain moiety contributes to ligand activity. This article describes a protocol that can be used to identify cleavage products generated during activation of Tgfß precursor proteins. RNA encoding Tgfß precursors are first microinjected into X. laevis embryos. The following day, cleavage products are collected from the blastocoele of gastrula stage embryos and analyzed on Western blots. This protocol can be completed relatively quickly, does not require expensive reagents and provides a source of concentrated Tgfß cleavage products under physiologic conditions.
???displayArticle.pubmedLink???
34369935
???displayArticle.pmcLink???PMC8717547 ???displayArticle.link???J Vis Exp ???displayArticle.grants???[+]
Birsoy,
XPACE4 is a localized pro-protein convertase required for mesoderm induction and the cleavage of specific TGFbeta proteins in Xenopus development.
2005, Pubmed,
Xenbase
Birsoy,
XPACE4 is a localized pro-protein convertase required for mesoderm induction and the cleavage of specific TGFbeta proteins in Xenopus development.
2005,
Pubmed
,
Xenbase
Constam,
Regulation of TGFβ and related signals by precursor processing.
2014,
Pubmed
Constam,
Regulation of bone morphogenetic protein activity by pro domains and proprotein convertases.
1999,
Pubmed
Cui,
The activity and signaling range of mature BMP-4 is regulated by sequential cleavage at two sites within the prodomain of the precursor.
2001,
Pubmed
,
Xenbase
Degnin,
Cleavages within the prodomain direct intracellular trafficking and degradation of mature bone morphogenetic protein-4.
2004,
Pubmed
,
Xenbase
Goldman,
Mutation of an upstream cleavage site in the BMP4 prodomain leads to tissue-specific loss of activity.
2006,
Pubmed
,
Xenbase
Gray,
Requirement for activin A and transforming growth factor--beta 1 pro-regions in homodimer assembly.
1990,
Pubmed
Harrison,
Prodomains regulate the synthesis, extracellular localisation and activity of TGF-β superfamily ligands.
2011,
Pubmed
Kim,
Proteolytic Activation of Bmps: Analysis of Cleavage in Xenopus Oocytes and Embryos.
2019,
Pubmed
,
Xenbase
Le Good,
Nodal stability determines signaling range.
2005,
Pubmed
Mimoto,
Manipulation of gene function in Xenopus laevis.
2011,
Pubmed
,
Xenbase
Nelsen,
Site-specific cleavage of BMP4 by furin, PC6, and PC7.
2009,
Pubmed
,
Xenbase
Nelsen,
Proprotein convertase genes in Xenopus development.
2005,
Pubmed
,
Xenbase
Neugebauer,
The prodomain of BMP4 is necessary and sufficient to generate stable BMP4/7 heterodimers with enhanced bioactivity in vivo.
2015,
Pubmed
,
Xenbase
Robertson,
Regulation of the Bioavailability of TGF-β and TGF-β-Related Proteins.
2016,
Pubmed
Sopory,
Regulation of bone morphogenetic protein-4 activity by sequence elements within the prodomain.
2006,
Pubmed
,
Xenbase
Tilak,
Simultaneous rather than ordered cleavage of two sites within the BMP4 prodomain leads to loss of ligand in mice.
2014,
Pubmed
,
Xenbase
Wang,
Structure and activation of pro-activin A.
2016,
Pubmed
Zhao,
Prodomain-growth factor swapping in the structure of pro-TGF-β1.
2018,
Pubmed
Zhou,
GDF6-CD99 Signaling Regulates Src and Ewing Sarcoma Growth.
2020,
Pubmed
