Fluorescence spectral properties of troponin C mutant F22W with one-, two-, and three-photon excitation
| Autor: | Joseph R. Lakowicz, Patrick K. Umeda, Ignacy Gryczynski, John Robinson, Henryk Malak, Herbert C. Cheung |
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| Rok vydání: | 1996 |
| Předmět: |
Photon
Protein Conformation Muscle Fibers Skeletal Analytical chemistry Biophysics Fluorescence Polarization Chick Embryo 02 engineering and technology Molecular physics Troponin C 03 medical and health sciences Animals Point Mutation Emission spectrum Muscle Skeletal Anisotropy 030304 developmental biology Photons 0303 health sciences Chemistry Tryptophan Hydrogen-Ion Concentration 021001 nanoscience & nanotechnology Fluorescence Recombinant Proteins Kinetics Spectrometry Fluorescence Mutagenesis Site-Directed 0210 nano-technology Chickens Excitation Fluorescence anisotropy Research Article |
| Zdroj: | Biophysical Journal. 71(6):3448-3453 |
| ISSN: | 0006-3495 |
| DOI: | 10.1016/s0006-3495(96)79540-1 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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