Defining elastic fiber interactions by molecular fishing: an affinity purification and mass spectrometry approach

Mol Cell Proteomics. 2009 Dec;8(12):2715-32. doi: 10.1074/mcp.M900008-MCP200. Epub 2009 Sep 15.


Deciphering interacting networks of the extracellular matrix is a major challenge. We describe an affinity purification and mass spectrometry strategy that has provided new insights into the molecular interactions of elastic fibers, essential extracellular assemblies that provide elastic recoil in dynamic tissues. Using cell culture models, we defined primary and secondary elastic fiber interaction networks by identifying molecular interactions with the elastic fiber molecules fibrillin-1, MAGP-1, fibulin-5, and lysyl oxidase. The sensitivity and validity of our method was confirmed by identification of known interactions with the bait proteins. Our study revealed novel extracellular protein interactions with elastic fiber molecules and delineated secondary interacting networks with fibronectin and heparan sulfate-associated molecules. This strategy is a novel approach to define the macromolecular interactions that sustain complex extracellular matrix assemblies and to gain insights into how they are integrated into their surrounding matrix.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium-Binding Proteins / metabolism
  • Chromatography, Affinity / methods*
  • Elastic Tissue / metabolism*
  • Fibrillin-1
  • Fibrillins
  • Heparin / metabolism
  • Humans
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Mass Spectrometry / methods*
  • Microfilament Proteins / chemistry
  • Microfilament Proteins / metabolism
  • Peptides / metabolism
  • Progranulins
  • Protein Binding
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Reproducibility of Results
  • Software


  • CLSTN1 protein, human
  • Calcium-Binding Proteins
  • FBN1 protein, human
  • Fibrillin-1
  • Fibrillins
  • GRN protein, human
  • Intercellular Signaling Peptides and Proteins
  • Microfilament Proteins
  • Peptides
  • Progranulins
  • Recombinant Proteins
  • Heparin