Abstract
Proprotein convertases (PCs) are secretory proteolytic enzymes that activate precursor proteins into biologically active forms by limited proteolysis at one or multiple internal sites. PCs are implicated in the processing of multiple protein precursors, including hormones, proteases, growth factors, angiogenic factors, and receptors. PCs have been linked recently to various pathologies such as Alzheimer’s disease, tumorigenesis, and infections. The zebrafish has emerged as an attractive model for studying the role of PCs not only in substrate production but also in development. Herein we describe methods that are used to characterize DNA sequences of PCs in zebrafish, as well as to evaluate the ontogeny and tissue distribution of their transcripts. We also provide information on the morpholino-mediated knockdown of proprotein convertases.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Seidah, N. G., and Chretien, M. (1999) Proprotein and prohormone convertases: A family of subtilases generating diverse bioactive polypeptides Brain Res 848, 45–62.
Seidah, N. G., Mowla, S. J., Hamelin, J., Mamarbachi, A. M., Benjannet, S., Toure, B. B., Basak, A., Munzer, J. S., Marcinkiewicz, J., Zhong, M., Barale, J. C., Lazure, C., Murphy, R. A., Chretien, M., and Marcinkiewicz, M. (1999) Mammalian subtilisin/kexin isozyme SKI-1: A widely expressed proprotein convertase with a unique cleavage specificity and cellular localization Proc Natl Acad Sci USA 96, 1321–6.
Seidah, N. G., Benjannet, S., Wickham, L., Marcinkiewicz, J., Jasmin, S. B., Stifani, S., Basak, A., Prat, A., and Chretien, M. (2003) The secretory proprotein convertase neural apoptosis-regulated convertase 1 (NARC-1): Liver regeneration and neuronal differentiation Proc Natl Acad Sci USA 100, 928–33.
Seidah, N. G., Mayer, G., Zaid, A., Rousselet, E., Nassoury, N., Poirier, S., Essalmani, R., and Prat, A. (2008) The activation and physiological functions of the proprotein convertases Int J Biochem Cell Biol 40, 1111–25.
Morash, M. G., MacDonald, A. B., Croll, R. P., and Anini, Y. (2009) Molecular cloning, ontogeny and tissue distribution of zebrafish (Danio rerio) prohormone convertases: PCSK1 and PCSK2 Gen Comp Endocrinol 162, 179–87.
Kudo, H., Liu, J., Jansen, E. J., Ozawa, A., Panula, P., Martens, G. J., and Lindberg, I. (2009) Identification of proSAAS homologs in lower vertebrates: Conservation of hydrophobic helices and convertase-inhibiting sequences Endocrinology 150, 1393–9.
Poirier, S., Prat, A., Marcinkiewicz, E., Paquin, J., Chitramuthu, B. P., Baranowski, D., Cadieux, B., Bennett, H. P., and Seidah, N. G. (2006) Implication of the proprotein convertase NARC-1/PCSK9 in the development of the nervous system J Neurochem 98, 838–50.
Walker, M. B., Miller, C. T., Coffin Talbot, J., Stock, D. W., and Kimmel, C. B. (2006) Zebrafish furin mutants reveal intricacies in regulating Endothelin1 signaling in craniofacial patterning Dev Biol 295, 194–205.
Nusslein-Volhard, C., and Dahm, R. (2002) Zebrafish: A Practical Approach. Oxford University Press, New York, NY and Cary, NC.
Bustin, S. A., Benes, V., Garson, J. A., Hellemans, J., Huggett, J., Kubista, M., Mueller, R., Nolan, T., Pfaffl, M. W., Shipley, G. L., Vandesompele, J., and Wittwer, C. T. (2009) The MIQE guidelines: Minimum information for publication of quantitative real-time PCR experiments Clin Chem 55, 611–22.
Acknowledgments
Research in Dr. Anini’s laboratory is supported by an operating grant from the Canadian Institutes of Health Research (CIHR) and an infrastructure grant from the Canada foundation of Innovation (CFI).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Morash, M.G., Soanes, K., Anini, Y. (2011). Prohormone Processing in Zebrafish. In: Mbikay, M., Seidah, N. (eds) Proprotein Convertases. Methods in Molecular Biology, vol 768. Humana Press. https://doi.org/10.1007/978-1-61779-204-5_14
Download citation
DOI: https://doi.org/10.1007/978-1-61779-204-5_14
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-203-8
Online ISBN: 978-1-61779-204-5
eBook Packages: Springer Protocols