Dynamics of Bacteriophage R17 Probed with a Long-Lifetime Ru(II) Metal-Ligand Complex |
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Authors: | Myung Sup Kim Jae Hui Kim Beng Whwa Son and Jung Sook Kang |
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Institution: | (1) Department of Oral Biochemistry and Molecular Biology, School of Dentistry, Pusan National University, Yangsan, 626-870, Korea;(2) Department of Chemistry, Pukyong National University, Busan, 608-737, Korea; |
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Abstract: | The metal-ligand complex, Ru(2,2′-bipyridine)2(4,4′-dicarboxy-2,2′-bipyridine)]2+ (RuBDc), was used as a spectroscopic probe for studying macromolecular dynamics. RuBDc is a very photostable probe that possesses
favorable photophysical properties including long lifetime, high quantum yield, large Stokes’ shift, and highly polarized
emission. To further show the usefulness of this luminophore for probing macromolecular dynamics, we examined the intensity
and anisotropy decays of RuBDc when conjugated to R17 bacteriophage using frequency-domain fluorometry with a blue light-emitting
diode (LED) as the modulated light source. The intensity decays were best fit by a sum of two exponentials, and we obtained
a longer mean lifetime at 4 °C (<τ> = 491.8 ns) as compared to that at 25 °C (<τ> = 435.1 ns). The anisotropy decay data showed
a single rotational correlation time, which is typical for a spherical molecule, and the results showed a longer rotational
correlation time at 4 °C (2,574.9 ns) than at 25 °C (2,070.1 ns). The use of RuBDc enabled us to measure the rotational correlation
time up to several microseconds. These results indicate that RuBDc has significant potential for studying hydrodynamics of
biological macromolecules. |
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