Proteolytic activation of Bmps: Analysis of cleavage in Xenopus oocytes and embryos

Hyung Seok Kim, Autumn McKnite, Jan Louise Christian

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Bone morphogenetic proteins (Bmps) are synthesized as inactive precursors that are cleaved to generate active ligands, along with prodomain fragments that can modulate growth factor activity. Here we provide three protocols that can be used to examine the process of proteolytic activation of Bmps. The first protocol describes how to generate radiolabeled Bmp precursor proteins in Xenopus oocytes and then analyze the time course of precursor cleavage by recombinant enzymes in vitro. The second protocol details how to analyze cleavage of radiolabeled precursor proteins in Xenopus oocytes over time using pulse-chase analysis and autoradiography. This protocol can also be used to analyze folding and cleavage of radiolabeled precursor proteins at steady state. Finally, the third protocol details methods for isolating Bmp cleavage products from the blastocoele of Xenopus embryos and then analyzing them on immunoblots.

LanguageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages115-133
Number of pages19
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1891
ISSN (Print)1064-3745

Fingerprint

Proprotein Convertases
Bone Morphogenetic Proteins
Protein Precursors
Xenopus laevis
Xenopus
Autoradiography
Xenopus Proteins
Oocytes
Embryonic Structures
Intercellular Signaling Peptides and Proteins
Ligands
Enzymes
caN protocol

Keywords

  • Autoradiography
  • Bone morphogenetic protein
  • Embryos
  • Immunoblot
  • Oocytes
  • Proprotein convertase
  • Proteolytic activation
  • Xenopus laevis

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Kim, H. S., McKnite, A., & Christian, J. L. (2019). Proteolytic activation of Bmps: Analysis of cleavage in Xenopus oocytes and embryos. In Methods in Molecular Biology (pp. 115-133). (Methods in Molecular Biology; Vol. 1891). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8904-1_9

Proteolytic activation of Bmps : Analysis of cleavage in Xenopus oocytes and embryos. / Kim, Hyung Seok; McKnite, Autumn; Christian, Jan Louise.

Methods in Molecular Biology. Humana Press Inc., 2019. p. 115-133 (Methods in Molecular Biology; Vol. 1891).

Research output: Chapter in Book/Report/Conference proceedingChapter

Kim, HS, McKnite, A & Christian, JL 2019, Proteolytic activation of Bmps: Analysis of cleavage in Xenopus oocytes and embryos. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1891, Humana Press Inc., pp. 115-133. https://doi.org/10.1007/978-1-4939-8904-1_9
Kim HS, McKnite A, Christian JL. Proteolytic activation of Bmps: Analysis of cleavage in Xenopus oocytes and embryos. In Methods in Molecular Biology. Humana Press Inc. 2019. p. 115-133. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-8904-1_9
Kim, Hyung Seok ; McKnite, Autumn ; Christian, Jan Louise. / Proteolytic activation of Bmps : Analysis of cleavage in Xenopus oocytes and embryos. Methods in Molecular Biology. Humana Press Inc., 2019. pp. 115-133 (Methods in Molecular Biology).
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