This title appears in the Scientific Report :
2013
Please use the identifier:
http://dx.doi.org/10.1039/c2pp25222h in citations.
Tryptophan fluorescence as a reporter for structural changes in photoactive yellow protein elicited by photo-activation.
Tryptophan fluorescence as a reporter for structural changes in photoactive yellow protein elicited by photo-activation.
Light-activation of photoactive yellow protein (PYP) is followed by a series of dynamical transitions in the structure of the protein. Tryptophan fluorescence is well-suited as a tool to study selected aspects of these. Using site-directed mutagenesis eight 'single-tryptophan' mutants of P...
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Personal Name(s): | Hospes, Marijke (Corresponding author) |
---|---|
Hendriks, Johnny / Hellingwerf, Klaas J | |
Contributing Institute: |
Zelluläre Biophysik; ICS-4 |
Published in: | Photochemical & photobiological sciences, 12 (2013) 3, S. 479 -488 |
Imprint: |
Cambridge
Royal Society of Chemistry
2013
|
PubMed ID: |
23183905 |
DOI: |
10.1039/c2pp25222h |
Document Type: |
Journal Article |
Research Program: |
Physics of the Cell |
Publikationsportal JuSER |
Light-activation of photoactive yellow protein (PYP) is followed by a series of dynamical transitions in the structure of the protein. Tryptophan fluorescence is well-suited as a tool to study selected aspects of these. Using site-directed mutagenesis eight 'single-tryptophan' mutants of PYP were made by replacement of either a tyrosine, phenylalanine or histidine residue by tryptophan, while simultaneously eliminating the endogenous W119. Surprisingly, only three of these eight mutants turn out to emit measurable tryptophan fluorescence: F6W/W119F, F96W/W119F and H108W/W119F. Significantly, all three show altered tryptophan fluorescence upon formation of the pB state. As F96 is located very close to the chromophore, the F96W/W119F mutant protein is particularly suitable for further studies on the dynamical changes of the polarity in the chromophore-binding pocket of PYP. Furthermore, WT PYP can be photo-activated by a UV photon via the highly conserved W119 and subsequent Förster resonance energy transfer. Placing a unique tryptophan residue elsewhere in the protein shows that position 119 is favoured for UV-activation of PYP. |