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Characterization of the EF-Hand Calcium-Binding Domains of Human Plastins

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Book cover Calcium-Binding Proteins of the EF-Hand Superfamily

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1929))

Abstract

The three human plastins (L-plastin, T-plastin, and I-plastin) are important regulatory Ca2+-binding proteins that belong to the family of actin-binding proteins. Plastins are involved in the regulation of the actin cytoskeleton as well as the cross-linking of actin filaments. In addition to four calponin-homology (CH) domains, all three plastins contain two N-terminal EF-hand Ca2+-binding motifs which together are homologous to a single lobe of the well-known calcium-regulatory protein calmodulin. This part of the protein allows for the regulation of the actin bundling activity in response to elevated calcium levels. In this protocol, we describe the purification of the EF-hand headpiece domains of all three plastins, as well as SPR studies, ITC studies, and NMR interaction studies with different peptides and calcium. In combination, these three experimental techniques provide detailed insights into a novel regulatory mechanism, involving the linker region between the EF-hand domain and the first CH domain of the plastins.

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Acknowledgments

This research is currently supported by a Discovery grant from the Natural Sciences and Engineering Research Council of Canada.

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Authors and Affiliations

  1. Biochemistry Research Group, Department of Biological Sciences, University of Calgary, Calgary, AB, Canada

    Oleg Khassan, Katharine V. Jensen, Andrew G. Woodman, Hans J. Vogel & Hiroaki Ishida

Authors
  1. Oleg Khassan
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  2. Katharine V. Jensen
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  3. Andrew G. Woodman
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  4. Hans J. Vogel
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  5. Hiroaki Ishida
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Corresponding author

Correspondence to Hans J. Vogel .

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Editors and Affiliations

  1. Division of Clinical Chemistry and Biochemistry, Department of Pediatrics, University of Zürich, Zürich, Switzerland

    Claus W. Heizmann

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Khassan, O., Jensen, K.V., Woodman, A.G., Vogel, H.J., Ishida, H. (2019). Characterization of the EF-Hand Calcium-Binding Domains of Human Plastins. In: Heizmann, C. (eds) Calcium-Binding Proteins of the EF-Hand Superfamily. Methods in Molecular Biology, vol 1929. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9030-6_16

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  • DOI: https://doi.org/10.1007/978-1-4939-9030-6_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9029-0

  • Online ISBN: 978-1-4939-9030-6

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