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Structural and lipid-binding characterization of human annexin A13a reveals strong differences with its long A13b isoform

  • Sara Fernández-Lizarbe , Emilio Lecona , Angélica Santiago-Gómez , Nieves Olmo , María Antonia Lizarbe and Javier Turnay EMAIL logo
Published/Copyright: September 26, 2016
Biological Chemistry
From the journal Biological Chemistry Volume 398 Issue 3

Abstract

Annexin A13 is the founder member of the vertebrate family of annexins, which are comprised of a tetrad of unique conserved domains responsible for calcium-dependent binding to membranes. Its expression is restricted to epithelial intestinal and kidney cells. Alternative splicing in the N-terminal region generates two isoforms, A13a and A13b, differing in a deletion of 41 residues in the former. We have confirmed the expression of both isoforms in human colon adenocarcinoma cells at the mRNA and protein levels. We have cloned, expressed, and purified human annexin A13a for the first time to analyze its structural characteristics. Its secondary structure and thermal stability differs greatly from the A13b isoform. The only tryptophan residue (Trp186) is buried in the protein core in the absence of calcium but is exposed to the solvent after calcium binding even though circular dichroism spectra are quite similar. Non-myristoylated annexin A13a binds in a calcium-dependent manner to acidic phospholipids but not to neutral or raft-like liposomes. Calcium requirements for binding to phosphatidylserine are around 6-fold lower than those required by the A13b isoform. This fact could account for the different subcellular localization of both annexins as binding to basolateral membranes seems to be calcium-dependent and myristoylation-independent.

Keywords: annexin; calcium binding; circular dichroism spectroscopy; fluorescence emission spectroscopy; phospholipid binding; thermal stability

Acknowledgments

We are thankful to the staff from the Genomics and Proteomics Center from the Complutense University of Madrid for their skillful assistance. This work was supported by the Ministerio de Ciencia e Innovación (MICINN), Spain (grant number BFU2008-04758).

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Supplemental Material:

The online version of this article (DOI: 10.1515/hsz-2016-0242) offers supplementary material, available to authorized users.

Received: 2016-6-30
Accepted: 2016-9-22
Published Online: 2016-9-26
Published in Print: 2017-3-1

©2017 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/hsz-2016-0242
Keywords for this article
annexin; calcium binding; circular dichroism spectroscopy; fluorescence emission spectroscopy; phospholipid binding; thermal stability

Articles in the same Issue

  1. Frontmatter
  2. Reviews
  3. Lysosomes in programmed cell death pathways: from initiators to amplifiers
  4. Mitochondrial carriers in inflammation induced by bacterial endotoxin and cytokines
  5. Recent insights into nitrite signaling processes in blood
  6. Research Articles/Short Communications
  7. Genes and Nucleic Acids
  8. Pokemon decreases the transcriptional activity of RARα in the absence of ligand
  9. Protein Structure and Function
  10. Comparison of enzymatic properties and small molecule inhibition of γ–glutamyltranspeptidases from pathogenic and commensal bacteria
  11. Structural and lipid-binding characterization of human annexin A13a reveals strong differences with its long A13b isoform
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  13. A role for the metalloprotease invadolysin in insulin signaling and adipogenesis
  14. Mirolysin, a LysargiNase from Tannerella forsythia, proteolytically inactivates the human cathelicidin, LL-37
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