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- Title
5-Fluorotryptophan as dual probe for ground-state heterogeneity and excited-state dynamics in apoflavodoxin
- Authors
Visser, N.V.; Westphal, A.H.; Nabuurs, S.M.; van Hoek, A.; van Mierlo, C.P.M.; Visser, A.J.W.G.; Broos, J.; van Amerongen, H.
- Abstract
Abstract: 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 19F 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 19F NMR and fluorescence spectroscopy.
- Subjects
TRYPTOPHAN; BACTERIAL proteins; AZOTOBACTER; BIOSYNTHESIS; FLUORESCENCE; NUCLEAR magnetic resonance; PROTEINS; DYNAMICS
- Publication
FEBS Letters, 2009, Vol 583, Issue 17, p2785
- ISSN
0014-5793
- Publication type
Article
- DOI
10.1016/j.febslet.2009.07.022