Fluorescence resonance energy transfer (FRET) as a method to calculate the dimerization strength of basic Helix-loop-Helix (bHLH) proteins

Victoria E. Centonze, Beth A. Firulli, Anthony Firulli

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Post-translational modifications such as phosphorylation play a vital role in the regulation of protein function. In our study of the basic Helix-loop-Helix (bHLH) transcription factor HAND1, we show that HAND1 is phosphorylated during the trophoblast giant cell differentiation on residues residing in Helix I of the bHLH domain. Our hypothesis is that these modifications result in changes in HAND1 dimerization affinities with other bHLH factors. To test this idea, we employed FRET to measure the protein-protein interactions of HAND1 and HAND1 point mutants in HEK293 cells using YFP and CFP fusion proteins and laser scanning confocal microscopy.

Original languageEnglish
Pages (from-to)78-82
Number of pages5
JournalBiological Procedures Online
Volume6
Issue number1
DOIs
StatePublished - May 12 2004

Fingerprint

Fluorescence Resonance Energy Transfer
Dimerization
Helix-Loop-Helix Motifs
Proteins
Basic Helix-Loop-Helix Transcription Factors
Phosphorylation
Confocal microscopy
HEK293 Cells
Trophoblasts
Giant Cells
Post Translational Protein Processing
Confocal Microscopy
Cell Differentiation
Fusion reactions
Scanning
Lasers

Keywords

  • Fluorescence resonance energy transfer
  • Phosphorylation
  • Transcription factors

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Fluorescence resonance energy transfer (FRET) as a method to calculate the dimerization strength of basic Helix-loop-Helix (bHLH) proteins. / Centonze, Victoria E.; Firulli, Beth A.; Firulli, Anthony.

In: Biological Procedures Online, Vol. 6, No. 1, 12.05.2004, p. 78-82.

Research output: Contribution to journalArticle

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