Wavelength Dependant Quenching of 2,5-Diphenyloxazole Fluorescence by Nucleotides |
| |
Authors: | N V KrishnaMurthy A R Reddy B Bhudevi |
| |
Institution: | (1) Jonaki, Labeled Biomolecules Laboratory, Regional Centre, Board of Radiation and Isotope Technology, Hyberabad, India;(2) Department of Chemistry, University College of Science, Osmania University, Hyberabad, 500 007, India |
| |
Abstract: | The quenching of 2,5-diphenyloxazole (PPO) fluorescence by nucleotides has been investigated by electronic absorption and
steady state fluorescence spectra. Five purine nucleotides AMP, ADP, ATP, GMP and dGMP, one pyrimidine nucleotide UMP and
one dinucleotide NAD have been employed in the present study. Electronic absorption studies indicate that there is no ground
state complexation between the nucleotides and PPO. The quenching of PPO fluorescence was investigated at two different wavelengths.
When excited at 304 nm, the λ
max of PPO, the fluorescence spectra of PPO is quenched following Stern–Volmer kinetics. The quenching ability of nucleotides
are in the order NAD > AMP > ADP > GMP > dGMP > UMP. The K
SV and k
q values obtained indicate that AMP is a better quencher of PPO fluorescence than GMP, which is contrary to commonly observed
pattern. The quenching is found to be dynamic in nature. However, when excited at 260 nm, the absorption maximum of the nucleotides,
the fluorescence intensity of PPO is reduced with increase in the concentration of the nucleotide. This is attributed to the
primary inner filter effect arising due to the absorption of the incident radiation by the nucleotides. Thus the inner filter
effect phenomenon can be employed to assay the non-fluorescent molecules by fluorimetry. |
| |
Keywords: | 2 5-Diphenyloxazole Fluorescence quenching Nucleotides Inner filter effect |
本文献已被 PubMed SpringerLink 等数据库收录! |
|