Probing the conformation of the fibronectin III1-2 domain by fluorescence resonance energy transfer

Nancy W. Karuri, Zong Lin, Hays S. Rye, Jean E. Schwarzbauer

Research output: Contribution to journalArticlepeer-review

25 Scopus citations


Fibronectin (FN) matrix is crucial for cell and tissue functions during embryonic development, wound healing, and oncogenesis. Assembly of FN matrix fibrils requires FN domains that mediate interactions with integrin receptors and with other FN molecules. In addition, regulation of FN matrix assembly depends on the first two FN type III modules, III1 and III2, which harbor FN-binding sites. We propose that interactions between these two modules sequester FN-binding sites in soluble FN and that these sites become exposed by FN conformational changes during assembly. To test the idea that III1-2 has a compact conformation, we constructed CIIIY, a conformational sensor of III1-2 based on fluorescent resonance energy transfer between cyan and yellow fluorescent proteins conjugated at its N and C termini.Wedemonstrate energy transfer in CIIIY and show that fluorescent resonance energy transfer was eliminated by proteolysis and by treatment with mild denaturants that disrupted intramolecular interactions between the two modules. We also show that mutations of key charged residues resulted in conformational changes that exposed binding sites for the N-terminal 70-kDa FN fragment. Collectively, these results support a conformation-dependent mechanism for the regulation of FN matrix assembly by III1-2.

Original languageEnglish (US)
Pages (from-to)3445-3452
Number of pages8
JournalJournal of Biological Chemistry
Issue number6
StatePublished - Feb 6 2009

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Biochemistry
  • Cell Biology


Dive into the research topics of 'Probing the conformation of the fibronectin III1-2 domain by fluorescence resonance energy transfer'. Together they form a unique fingerprint.

Cite this