5-fluorotryptophan as dual probe for ground-state heterogeneity and excited-state dynamics in apoflavodoxin

N.V. Visser, A.H. Westphal, S.M. Nabuurs, A. van Hoek, C.P.M. van Mierlo, A.J.W.G. Visser, J. Broos, H. van Amerongen

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12 Citations (Scopus)


The apoflavodoxin protein from Azotobacter vinelandii harboring three tryptophan (Trp) residues, was biosynthetically labeled with 5-fluorotryptophan (5-FTrp). 5-FTrp has the advantage that chemical differences in its microenvironment can be sensitively visualized via F-19 NMR. Moreover, it shows simpler fluorescence decay kinetics. The occurrence of FRET was earlier observed via the fluorescence anisotropy decay of WT apoflavodoxin and the anisotropy decay parameters are in excellent agreement with distances between and relative orientations of all Trp residues. The anisotropy decay in 5-FTrp apoflavodoxin demonstrates that the distances and orientations are identical for this protein. This work demonstrates the added value of replacing Trp by 5-FTrp to study structural features of proteins via F-19 NMR and fluorescence spectroscopy.
Original languageEnglish
Pages (from-to)2785-2788
Number of pages3
JournalFEBS Letters
Issue number17
Publication statusPublished - 3 Sept 2009


  • fluorescence anisotropy
  • F-19 NMR
  • picosecond fluorescence
  • protein folding
  • protein stability
  • azotobacter-vinelandii
  • fluorescence decay
  • tryptophan
  • proteins
  • kinetics
  • pathway


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