Fluorescence spectral properties of troponin C mutant F22W with one-, two-, and three-photon excitation

Biophysical Journal
I GryczynskiP K Umeda

Abstract

We report the first measurements of protein fluorescence with three-photon excitation, using a mutant of troponin C (TnC) that contains a single tryptophan residue F22W. From the emission intensity dependence on laser power we determine that TnC F22W displays one-, two-, and three-photon excitation at 285, 570, and 855 nm, respectively. The emission spectra and intensity decays are identical for one-, two-, or three-photon excitation. The steady-state and time 0 anisotropies are distinct for each mode of excitation, but the correlation times were the same, suggesting that three-photon excitation of proteins can be accomplished without significant effects of the locally intense illumination. The excitation anisotropy spectrum from 830 to 900 nm displays only negative values, suggesting dominant excitation via the 1Lb state of tryptophan from 830 to 900 nm.

References

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Citations

Apr 17, 2001·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·J R LakowiczI Gryczynski
Jun 17, 1999·Biophysical Chemistry·I GryczynskiJ R Lakowicz
Aug 7, 2013·Nano Letters·Christopher M GallowayRomain Quidant
Sep 1, 1996·Journal of Fluorescence·I GryczynskiJ R Lakowicz
May 7, 2015·Der Hautarzt; Zeitschrift für Dermatologie, Venerologie, und verwandte Gebiete·M ZiegerM Kaatz
Apr 5, 1997·Analytical Biochemistry·I GryczynskiJ R Lakowicz
Jun 6, 2000·Journal of Biomolecular Screening·J R LakowiczZ Gryczynski
Feb 1, 2002·The Journal of Physical Chemistry. a·Ignacy GryczynskiJoseph R Lakowicz
Feb 16, 2006·Biochemistry·Sebastian BreuerMatthias Geyer

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