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Withaferin A binds covalently to the N-terminal domain of annexin A2

  • Gabriel Ozorowski , Christopher M. Ryan , Julian P. Whitelegge and Hartmut Luecke EMAIL logo
Published/Copyright: September 8, 2012
Biological Chemistry
From the journal Biological Chemistry Volume 393 Issue 10

Abstract

Annexin A2 (AnxA2), a 38-kDa member of the Ca2+-binding annexin family, has been implicated in numerous cancer pathways. Withaferin A (WithfA), a natural plant compound, has been reported previously to bind covalently to Cys133 of the AnxA2 core domain leading to a reduction of the invasive capabilities of cancer cells by altering their cytoskeleton. We show here that AnxA2 has an inhibitory effect on actin polymerization, and a modification with WithfA significantly increases this inhibitory role of AnxA2. Using mass spectrometry and single-site mutants, we localized the WithfA-AnxA2 interaction to the N-terminal domain of AnxA2 where WithfA binds covalently to Cys9. Whereas binding to F-actin filaments has been mapped to the C terminus of AnxA2, our results suggest that the N-terminal domain modified by WithfA may also play a role in the AnxA2-actin interaction. The binding of WithfA may regulate the AnxA2-mediated actin dynamics in two distinct ways: (i) the increase of F-actin bundling activity by the Anx2/p11 heterotetramer and (ii) the decrease of actin polymerization as a result of the increased affinity of AnxA2 to the barbed end of actin microfilaments. We demonstrate the susceptibility of Cys9 of AnxA2 to chemical modifications and exclude Cys133 as a binding site for WithfA.

Keywords: actin polymerization; annexin A2, covalent modification; mass spectrometry; Robert Huber; withaferin A
Corresponding author: Hartmut Luecke, Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697-3900, USA
Received: 2012-4-19
Accepted: 2012-7-12
Published Online: 2012-09-08
Published in Print: 2012-10-01

©2012 by Walter de Gruyter Berlin Boston

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https://doi.org/10.1515/hsz-2012-0184
Keywords for this article
actin polymerization; annexin A2, covalent modification; mass spectrometry; Robert Huber; withaferin A

Articles in the same Issue

  1. Masthead
  2. Masthead
  3. Guest Editorial
  4. No end in sight: the development of protein crystallography in Martinsried
  5. Highlight: No End in Sight: The Development of Protein Crystallography in Martinsried
  6. Functional and structural insights into astacin metallopeptidases
  7. β-Trefoil inhibitors – from the work of Kunitz onward
  8. Structural basis of the TAL effector–DNA interaction
  9. Nature’s way of handling a greenhouse gas: the copper-sulfur cluster of purple nitrous oxide reductase
  10. Rustless translation
  11. Structural basis of [NiFe] hydrogenase maturation by Hyp proteins
  12. Covalent and non-covalent reversible proteasome inhibition
  13. Structural origins of AGC protein kinase inhibitor selectivities: PKA as a drug discovery tool
  14. Structural analysis of the C-terminal domain of the spliceosomal helicase Prp22
  15. N-terminal acetylation of annexin A2 is required for S100A10 binding
  16. Withaferin A binds covalently to the N-terminal domain of annexin A2
  17. PhoB transcriptional activator binds hierarchically to pho box promoters
  18. Research Articles/Short Communications
  19. Protein Structure and Function
  20. Biochemical and functional characterization of human phospholipid scramblase 4 (hPLSCR4)
  21. Cell Biology and Signaling
  22. Dual-specificity phosphatases are targets of the Wnt/β-catenin pathway and candidate mediators of β-catenin/Ras signaling interactions
  23. Proteolysis
  24. Inhibition of Aeromonas sobria serine protease (ASP) by α2-macroglobulin
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