Fluorescence spectral properties of troponin C mutant F22W with one-, two-, and three-photon excitation.
AUTOR(ES)
Gryczynski, I
RESUMO
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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1233832Documentos Relacionados
- Time-Resolved Fluorescence Spectroscopy and Imaging of DNA Labeled with DAPI and Hoechst 33342 Using Three-Photon Excitation
- Three-photon induced fluorescence of the calcium probe Indo-1.
- Changes in fluorescence, turbidity, and birefringence associated with nerve excitation.
- Simultaneous two-photon excitation of distinct labels for dual-color fluorescence crosscorrelation analysis
- Two-photon excitation microscopy of tryptophan-containing proteins