Structural interlock between ligand-binding site and stalk-like region of β1 integrin revealed by a monoclonal antibody recognizing conformation-dependent epitope

Junichi Takagi, Takashi Isobe, Yoshikazu Takada, Yuji Saito

Research output: Contribution to journalArticlepeer-review

31 Citations (Scopus)

Abstract

Integrin activation and sebsequent ligand binding to it are regulated by intracellular mechanisms called inside-out signaling, which are not fully understood and are accompanied by dynamic structural changes of the integrin molecule itself. A monoclonal antibody recognizing a conformation-dependent epitope on human β1 integrin was produced and characterized in detail. This antibody, AG89, reacted with human integrin β1 chain regardless of the α subunit. AG89 can recognize resting state β1 integrin on the cells, but the reactivity is increased ~ 2-fold upon integrin activation by activating anti-β1 antibodies and ~ 3-fold by Mn2+. Furthermore, occupation of the ligand-binding pocket by a soluble ligand (RGD peptide for αvβ1 and CS-1 peptide for α4β1) resulted in maximum binding of AG89, indicating that the epitope for AG89 is exposed during the conformational changes of β1 integrin upon activation/ligation. Epitope mapping by using interspecies chimeric β1 revealed that the epitope for AG89 lies within residues 426-587, which corresponds to the cysteine-rich repeat structure located in the middle of the β1 chain. The fact that binding of AG89 itself could activate the resting β1 integrin indicates that exposure of the AG89 epitope in the membrane-proximal stalk-like domain and 'opening' of the ligand-binding pocket at the outermost domain are physically linked. We propose that the integrin 'signaling' is mediated by this direct physical transduction of conformational information along the integrin molecule.

Original languageEnglish
Pages (from-to)914-921
Number of pages8
JournalJournal of Biochemistry
Volume121
Issue number5
Publication statusPublished - May 1997
Externally publishedYes

Keywords

  • Cell adhesion
  • Inside-out signaling
  • Integrin
  • Ligand binding
  • Monoclonal antibody

ASJC Scopus subject areas

  • Biochemistry

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