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Expression and characterization of a dictyostelium discoideum annexin

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Abstract

The annexins are calcium-dependent phospholipid-binding proteins. Recently the gene encoding the homologue of a mammalian annexin has been identified inDictyostelium discoideum. Analysis of cDNA and genomic clones showed that the transcript forDictyostelium annexin is alternatively spliced (Greenwood, M. and Tsang, A. (1991) Biochim. Biophys. Acta 1088, 429–432; Döring, V., Schleicher, M and Noegel, A. (1991) J. Biol. Chem. 266, 17509–17515). Here, we showed that theDictyostelium annexin DNA hybridized to two populations of transcripts. We used a recombinant annexin polypeptide to raise polyclonal antibody. Immunoblot analysis revealed that the antibody recognized two polypeptides of 48 kDa and 54 kDa in developingD. discoideum cells. The molecular sizes of these polypeptides correspond well with the expected sizes of the alternatively spliced products. The 48-kDa and 54-kDa polypeptides were purified by isoelectric focusing to more than 70% homogeneity. The partially purified proteins were found to associate with phosphatidylserine vesicles in a calcium-dependent manner. These results suggest that the 48- and 54-kDa polypeptides are the products of alternative splicing of the annexin transcripts. During development the two polypeptides accumulate at different rates to about 60 times the level detected in vegetative cells. On the other hand, RNA blot analysis showed that the level of the annexin transcripts in multicellular aggregates was about 5 times that of vegetative cells.

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Author notes

  1. Michael Greenwood

    Present address: Department of Medicine, McGill University, Royal Victoria Hospital, 687 Pine Avenue W., H3A 1A1, Montreal, Quebec, Canada

Authors and Affiliations

  1. Department of Biology, Concordia University, 1455 de Maisonneuve Blvd. W., H3G 1M8, Montreal, Quebec, Canada

    Claire Bonfils, Michael Greenwood & Adrian Tsang

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  1. Claire Bonfils
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  2. Michael Greenwood
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  3. Adrian Tsang
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Bonfils, C., Greenwood, M. & Tsang, A. Expression and characterization of a dictyostelium discoideum annexin. Mol Cell Biochem 139, 159–166 (1994). https://doi.org/10.1007/BF01081739

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  • DOI: https://doi.org/10.1007/BF01081739

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