Studies of receptor tyrosine kinase transmembrane domain interactions: The EmEx-FRET method

Mikhail Merzlyakov, Lirong Chen, Kalina A Hristova

Research output: Contribution to journalArticle

Abstract

The energetics of transmembrane (TM) helix dimerization in membranes and the thermodynamic principles behind receptor tyrosine kinase (RTK) TM domain interactions during signal transduction can be studied using Förster resonance energy transfer (FRET). For instance, FRET studies have yielded the stabilities of wild-type fibroblast growth factor receptor 3 (FGFR3) TM domains and two FGFR3 pathogenic mutants, Ala391Glu and Gly380Arg, in the native bilayer environment. To further our understanding of the molecular mechanisms of deregulated FGFR3 signaling underlying different pathologies, we determined the effect of the Gly382Asp FGFR3 mutation, identified in a multiple myeloma cell line, on the energetics of FGFR3 TM domain dimerization. We measured dimerization energetics using a novel FRET acquisition and processing method, termed "emission-excitation FRET (EmEx-FRET)," which improves the precision of thermodynamic measurements of TM helix association. The EmEx-FRET method, verified here by analyzing previously published data for wild-type FGFR3 TM domain, should have broad utility in studies of protein interactions, particularly in cases when the concentrations of fluorophore-tagged molecules cannot be controlled.

Original languageEnglish (US)
Pages (from-to)93-103
Number of pages11
JournalJournal of Membrane Biology
Volume215
Issue number2-3
DOIs
StatePublished - Feb 2007

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Receptor, Fibroblast Growth Factor, Type 3
Receptor Protein-Tyrosine Kinases
Energy Transfer
Dimerization
Thermodynamics
Multiple Myeloma
Signal Transduction
Pathology
Cell Line
Mutation
Membranes

Keywords

  • FRET
  • Receptor tyrosine kinase
  • Transmembrane domain

ASJC Scopus subject areas

  • Biophysics
  • Physiology
  • Cell Biology

Cite this

Studies of receptor tyrosine kinase transmembrane domain interactions : The EmEx-FRET method. / Merzlyakov, Mikhail; Chen, Lirong; Hristova, Kalina A.

In: Journal of Membrane Biology, Vol. 215, No. 2-3, 02.2007, p. 93-103.

Research output: Contribution to journalArticle

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