Characterization of human p33/41 (annexin VI), a Ca2+ dependent carbohydrate-binding protein with monoclonal anti-annexin IV antibodies, AS11 and AS17

Ayano Satoh, Eiji Takayama, Kyoko Kojima, Haruko Ogawa, Yoshimoto Katsura, Tatsuo Kina, Isamu Matsumoto

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

p33/41 (annexin IV) is a member of the family of Ca2+-dependent phospholipid binding proteins known as annexins. We previously described that bovine kidney p33/41 (annexin IV) has Ca2+-dependent carbohydrate binding activity. In this study, we purified human p33/41 (annexin IV) from the HT29, human colon adenocarcinoma cell line, as well as the bovine kidney annexin by affinity chromatography. Then, we prepared recombinant human p33/41 (annexin IV) expressed in Escherichia coli. The apparent size and the Ca2+-dependent carbohydrate binding properties of purified recombinant p33/41 (annexin IV) were indistinguishable from those of the bovine kidney protein. We also performed inhibition assays of carbohydrate binding and of phosphatidylserine/phosphatidylcholine liposome binding of recombinant p33/41 (annexin IV) with anti-p33/41 monoclonal antibodies (AS11 and AS17). We determined the epitopes recognized by the monoclonal antibodies b) Western blot analysis using deleted-recombinant p33/41 (annexin IV). The monoclonal antibodies recognized domain 1 and/or 2 of p33/41 (annexin IV). The results of the inhibition assays and the determination of the epitope showed that a carbohydrate binding site is located at domains 3 and 4 of p33/41 (annexin IV) and on the cell surface.

Original languageEnglish
Pages (from-to)224-229
Number of pages6
JournalBiological and Pharmaceutical Bulletin
Volume20
Issue number3
DOIs
Publication statusPublished - Mar 1997
Externally publishedYes

Keywords

  • annexin
  • lectin
  • monoclonal antibody

ASJC Scopus subject areas

  • Pharmacology
  • Pharmaceutical Science

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