A proteomic approach reveals integrin activation state-dependent control of microtubule cortical targeting

Nat Commun. 2015 Jan 22:6:6135. doi: 10.1038/ncomms7135.

Abstract

Integrin activation, which is regulated by allosteric changes in receptor conformation, enables cellular responses to the chemical, mechanical and topological features of the extracellular microenvironment. A global view of how activation state converts the molecular composition of the region proximal to integrins into functional readouts is, however, lacking. Here, using conformation-specific monoclonal antibodies, we report the isolation of integrin activation state-dependent complexes and their characterization by mass spectrometry. Quantitative comparisons, integrating network, clustering, pathway and image analyses, define multiple functional protein modules enriched in a conformation-specific manner. Notably, active integrin complexes are specifically enriched for proteins associated with microtubule-based functions. Visualization of microtubules on micropatterned surfaces and live cell imaging demonstrate that active integrins establish an environment that stabilizes microtubules at the cell periphery. These data provide a resource for the interrogation of the global molecular connections that link integrin activation to adhesion signalling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allosteric Site
  • Antibodies, Monoclonal / chemistry
  • Cerebral Cortex / metabolism
  • Cluster Analysis
  • Dimethylpolysiloxanes / chemistry
  • Fibroblasts / metabolism
  • Foreskin / metabolism
  • Green Fluorescent Proteins / metabolism
  • HeLa Cells
  • Humans
  • Integrin beta1 / metabolism
  • Integrins / metabolism*
  • K562 Cells
  • Male
  • Mass Spectrometry
  • Microscopy, Fluorescence
  • Microtubules / metabolism*
  • Protein Binding
  • Protein Conformation
  • Proteome
  • Proteomics / methods*
  • Signal Transduction

Substances

  • Antibodies, Monoclonal
  • Dimethylpolysiloxanes
  • Integrin beta1
  • Integrins
  • Proteome
  • Green Fluorescent Proteins
  • baysilon