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Purification and Assays of Tachylectin-2

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Lectin Purification and Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2132))

Abstract

Tachylectin-2, a 27-kDa protein consisting of a five-bladed β-propeller structure, is purified by three steps of chromatography, including dextran sulfate-Sepharose CL-6B, CM-Sepharose CL-6B, and Mono S. Three isolectins of tachylectin-2 including tachylectin-2a, -2b, and -2c are purified. These isolectins exhibit hemagglutinating activity against human A-type erythrocytes in a Ca2+-independent manner with tachylectin-2b showing the highest activity. Tachylectin-2b specifically agglutinates Staphylococcus saprophyticus KD. The tachylectin-2b-mediated hemagglutination is inhibited in the presence of GlcNAc and GalNAc. The association constants for GlcNAc and GalNAc are Ka = 1.95 × 104 M−1 and Ka = 1.11 × 103 M−1, respectively. Ultracentrifugation analysis shows that tachylectin-2b is present in monomer form in solution.

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Authors and Affiliations

  1. Department of Biology, Faculty of Science, Kyushu University, Fukuoka, Japan

    Shun-ichiro Kawabata & Toshio Shibata

Authors
  1. Shun-ichiro Kawabata
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  2. Toshio Shibata
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Correspondence to Shun-ichiro Kawabata .

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Editors and Affiliations

  1. Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan

    Jun Hirabayashi

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Kawabata, Si., Shibata, T. (2020). Purification and Assays of Tachylectin-2. In: Hirabayashi, J. (eds) Lectin Purification and Analysis. Methods in Molecular Biology, vol 2132. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0430-4_30

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  • DOI: https://doi.org/10.1007/978-1-0716-0430-4_30

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0429-8

  • Online ISBN: 978-1-0716-0430-4

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