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

Molecular & cellular proteomics 8, 2715-2732 (2009). doi:10.1074/mcp.M900008-MCP200
Deciphering interacting networks of the extracellular ma-trix is a major challenge. We describe an affinity purifica-tion and mass spectrometry strategy that has providednew insights into the molecular interactions of elasticfibers, essential extracellular assemblies that provideelastic recoil in dynamic tissues. Using cell culture mod-els, we defined primary and secondary elastic fiber inter-action networks by identifying molecular interactions withthe elastic fiber molecules fibrillin-1, MAGP-1, fibulin-5,and lysyl oxidase. The sensitivity and validity of ourmethod was confirmed by identification of known inter-actions with the bait proteins. Our study revealed novelextracellular protein interactions with elastic fiber mole-cules and delineated secondary interacting networks withfibronectin and heparan sulfate-associated molecules.This strategy is a novel approach to define the macromo-lecular interactions that sustain complex extracellularmatrix assemblies and to gain insights into how they areintegrated into their surrounding matrix
Published by The American Society for Biochemistry and Molecular Biology, Bethesda, Md.