Structural requirements for activation in αIIbβ3 integrin

Tetsuji Kamata, Makoto Handa, Sonomi Ito, Yukiko Sato, Toshimitsu Ohtani, Yohko Kawai, Yasuo Ikeda, Sadakazu Aiso

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Integrins are postulated to undergo structural rearrangement from a low affinity bent conformer to a high affinity extended conformer upon activation. However, some reports have shown that a bent conformer is capable of binding a ligand, whereas another report has shown that integrin extension does not absolutely lead to activation. To clarify whether integrin affinity is indeed regulated by the so-called switchblade-like movement, we have engineered a series of mutant αIIbβ3 integrins that are constrained specifically in either a bent or an extended conformation. These mutant αIIbβ3 integrins were expressed in mammalian cells, and fibrinogen binding to these cells was examined. The bent integrins were created through the introduction of artificial disulfide bridges in the β-head/β-tail interface. Cells expressing bent integrins all failed to bind fibrinogen unless pretreated with DTT to disrupt the disulfide bridges. The extended integrins were created by introducing N-glycosylation sites in amino acid residues located close to the α-genu, where the integrin legs fold backward. Among these mutants, activation was maximized in one integrin with an N-glycosylation site located behind the α-genu. This extension-induced activation was completely blocked when the swing-out of the hybrid domain was prevented. These results suggest that the bent and extended conformers represent low affinity and high affinity conformers, respectively, and that extension-induced activation depends on the swing-out of the hybrid domain. Taken together, these results are consistent with the current hypothesis that integrin affinity is regulated by the switchblade-like movement of the integrin legs.

Original languageEnglish
Pages (from-to)38428-38437
Number of pages10
JournalJournal of Biological Chemistry
Volume285
Issue number49
DOIs
Publication statusPublished - 2010 Dec 3

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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  • Cite this

    Kamata, T., Handa, M., Ito, S., Sato, Y., Ohtani, T., Kawai, Y., Ikeda, Y., & Aiso, S. (2010). Structural requirements for activation in αIIbβ3 integrin. Journal of Biological Chemistry, 285(49), 38428-38437. https://doi.org/10.1074/jbc.M110.139667