Fluorescence of horse liver alcohol dehydrogenase using one- and two-photon excitation |
| |
| Authors: | Joseph R Lakowicz Borys Kierdaszuk Ignacy Gryczynski Henryk Malak |
| |
| Institution: | (1) Center for Fluorescence Spectroscopy, Department of Biological Chemistry, University of Maryland School of Medicine, 108 North Greene Street, 21201 Baltimore, Maryland;(2) Department of Biophysics, University of Warsaw, 93 Zwirki i Wigury Street, PL-02089 Warsaw, Poland |
| |
| Abstract: | We examined the steady-state and time-resolved emission of liver alcohol dehydrogenase resulting from one-photon and two-photon excitation. Previous studies with one-photon excitation revealed that the two nonidentical tryptophan residues display different emission spectra and decay times. The use of two-photon excitation resulted in similar emission spectra, multiexponential intensity decays, time-resolved emission spectra, and anisotropy decays as was observed for one-photon excitation. These results suggest that both nonidentical tryptophan residues are excited to a similar extent for one- and two-photon excitation. However, the limiting anisotropy (r
0) with two-photon excitation from 585 to 610 nm is below 0.1 and appears distinct from that observed previously forN-acetyl-l-tryptophanamide.Abbreviations LADH
liver alcohol dehydrogenase
-  -NAD+
-nicotinamide adenine dinucleotide
- OPE
one-photon excitation
- OPIF
one-photon induced fluorescence
- TPE
two-photon excitation
- TCSPC
time-correlated single photon counting
- TPIF
two-photon induced fluorescence |
| |
| Keywords: | Fluorescence anisotropy one-photon excitation two-photon excitation anisotropy spectra horse liver alcohol dehydrogenase time-resolved fluorescence proteins tryptophan |
| 本文献已被 SpringerLink 等数据库收录！ |
|
