Annexin Lectins: Ca2+-Dependent Heparin-Binding Activity, Phosphatidylserine-Binding Activity, and Anticoagulant Activity

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


Among numerous heparin-binding proteins identified in animal tissues and body fluids, annexins are unique because their activies depend on their Ca2+ binding. Annexins are known to have other Ca2+-dependent activities. For example, they bind to phosphatidylserine in the plasma membrane, and some of them exhibit potent anticoagulant activity. This chapter describes three protocols that measure the Ca2+-dependent activities using recombinant annexins: solid-phase heparin-binding assay using bovine serum albumin-conjugated heparin, solid-phase phosphatidylserine-binding assay, and plasma coagulation inhibition assay.

Key words

Annexins Anticoagulant activity Glycosaminoglycans Heparin-binding activity Phosphatidylserine-binding activity 


  1. 1.
    Srikrishna G, Panneerselvam K, Westphal V et al (2001) Two proteins modulating transendothelial migration of leukocytes recognize novel carboxylated glycans on endothelial cells. J Immunol 166:4678–4688CrossRefGoogle Scholar
  2. 2.
    Gao C-X, Honke K, Taniguchi N (2004) Carbohydrate binding activity of annexin V toward a bisecting N-acetylglucosamine. Methods Enzymol 363:34–47CrossRefGoogle Scholar
  3. 3.
    Gao-Uozumi C-X, Uozumi N, Miyoshi E et al (2000) A novel carbohydrate binding activity of annexin V toward a bisecting N-acetylglucosamine. Glycobiology 10:1209–1216CrossRefGoogle Scholar
  4. 4.
    Kojima K, Ogawa HK, Seno N et al (1992) Carbohydrate-binding proteins in bovine kidney have consensus amino acid sequences of annexin family proteins. J Biol Chem 267:20536–20539PubMedGoogle Scholar
  5. 5.
    Ishitsuka R, Kojima K, Utsumi H et al (1998) Glycosaminoglycan binding properties of annexin IV, V, and VI. J Biol Chem 273:9935–9941CrossRefGoogle Scholar
  6. 6.
    Shao C, Zhang F, Kemp MM et al (2006) Crystallographic analysis of calcium-dependent heparin binding to annexin A2. J Biol Chem 281:31689–31695CrossRefGoogle Scholar
  7. 7.
    Capila I, Hernaiz MJ, Mo YD et al (2001) Annexin V-heparin oligosaccharide complex suggests heparan sulfate-mediated assembly on cell surfaces. Structure 9:57–64CrossRefGoogle Scholar
  8. 8.
    Bitter T, Muir HM (1962) A modified uronic acid carbazole reaction. Anal Biochem 4:330–334CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Graduate School of Humanities and SciencesOchanomizu UniversityTokyoJapan
  2. 2.Natural Science Division, Faculty of Core ResearchOchanomizu UniversityTokyoJapan

Personalised recommendations