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Bone Morphogenetic Proteins pp 115–133Cite as

Proteolytic Activation of Bmps: Analysis of Cleavage in Xenopus Oocytes and Embryos

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1891))

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.

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Author notes

  1. Hyung-Seok Kim and Autumn McKnite are contributed equally to this work.

Authors and Affiliations

  1. Division of Hematology and Hematologic Malignancies, Department of Neurobiology, Anatomy and Internal Medicine, University of Utah School of Medicine, Salt Lake City, UT, USA

    Hyung-Seok Kim, Autumn McKnite & Jan L. Christian

Authors
  1. Hyung-Seok Kim
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  2. Autumn McKnite
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  3. Jan L. Christian
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Corresponding author

Correspondence to Jan L. Christian .

Editor information

Editors and Affiliations

  1. New Jersey Medical School, Rutgers Biomedical and Health Sciences, Newark, NJ, USA

    Melissa B. Rogers

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Kim, HS., McKnite, A., Christian, J.L. (2019). Proteolytic Activation of Bmps: Analysis of Cleavage in Xenopus Oocytes and Embryos. In: Rogers, M. (eds) Bone Morphogenetic Proteins. Methods in Molecular Biology, vol 1891. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8904-1_9

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  • DOI: https://doi.org/10.1007/978-1-4939-8904-1_9

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8903-4

  • Online ISBN: 978-1-4939-8904-1

  • eBook Packages: Springer Protocols

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