Longistatin, an EF-Hand Ca2+-Binding Protein from Vector Tick: Identification, Purification, and Characterization

  • Anisuzzaman
  • M. Khyrul Islam
  • M. Abdul Alim
  • Naotoshi TsujiEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 963)


EF-hand Ca2+-binding motif, a structural component of the EF-hand protein, functions as a calcium sensor and/or buffer in the cytosol of the cell. However, in a few exceptional cases, the EF-hand proteins are secreted from cells and play crucial roles extracellularly. We have identified longistatin, an EF-hand Ca2+-binding protein, from the salivary glands of the tick, Haemaphysalis longicornis. Longistatin possesses an N-terminal sequence of unknown structure and two EF-hand motifs in the C-terminus, which conserve a calmodulin-like canonical structure. Longistatin shows distinct changes in its migration during electrophoresis through SDS-PAGE gel containing calcium or ethylenediaminetetraacetic acid (EDTA). Both recombinant and endogenous forms of longistatin can be stained with rutheninum red, demonstrating that longistatin is a Ca2+-binding protein.

Key words

Ticks Haemaphysalis longicornis Longistatin EF-hand motif Ca2+-binding protein 


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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Anisuzzaman
    • 1
  • M. Khyrul Islam
    • 2
  • M. Abdul Alim
    • 2
  • Naotoshi Tsuji
    • 1
    Email author
  1. 1.Department of Global Agricultural Sciences, Graduate School of Agricultural and Life SciencesThe University of TokyoTokyoJapan
  2. 2.National Institute of Animal Health, National Agricultural and Food Research OrganizationIbarakiJapan

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