The difference spectrum of the quercetin--DNA complex versus quercetin alone was characterized by a peak at 395 nm. An increase in the magnitude of difference spectrum was seen with increased ionic strength. Spectrophotometric changes in absorbance and fluorescence of quercetin showed that ethidium bromide is able to displace quercetin from the quercetin--DNA complex. These results indicate that the binding of quercetin to DNA does not involve electrostatic interactions but may be intercalative in nature. Experiments using DNase I footprinting technique showed that the flavonoid does not possess any preferred sites of binding in DNA. Strand scission in DNA by the quercetin--Cu(II) system gave a generally uniform cutting pattern of internucleotide bonds. This led to the observation that the quercetin--Cu(II) cleavage reaction has the potential of being used as preferred DNA footprinting reagent.
The objective of this study was to establish relationship between manganese-induced toxicity and antioxidant system response in Brassica juncea plants and also to investigate whether brassinosteroids activate antioxidant system to confer tolerance to the plants affected with manganese induced oxidative stress. Brassica juncea plants were administered with 3, 6, or 9 mM manganese at 10-day stage for 3 days. At 31-day stage, the seedlings were sprayed with deionized water (control) or 10(-8) M of 24-epibrassinolide, and plants were harvested at 45-day stage to assess growth, leaf gas-exchange traits, and biochemical parameters. The manganese treatments diminished growth along with photosynthetic attributes and carbonic anhydrase activity in the concentration-dependent manner, whereas it enhanced lipid peroxidation, electrolyte leakage, accumulation of H2O2 as well as proline, and various antioxidant enzymes in the leaves of Brassica juncea which were more pronounced at higher concentrations of manganese. However, the follow-up application of 24-epibrassinolide to the manganese stressed plants improved growth, water relations, and photosynthesis and further enhanced the various antioxidant enzymes viz. catalase, peroxidase, and superoxide dismutase and content of proline. The elevated level of antioxidant enzymes as well as proline could have conferred tolerance to the manganese-stressed plants resulting in improved growth and photosynthetic attributes.
The flavonoid, quercetin, is known to bind to DNA and, in the presence of Cu(II) and other ions, causes fragmentation of the molecule. We examined whether quercetin might bind to protein and cause similar fragmentation. By using UV spectroscopic and fluorescence quenching experiments we show that quercetin binds to bovine serum albumin and that the complex does, in the presence of Cu(II), lead to fragmentation of the protein. The binding involves binding to tryptophan residues in the albumin. The reaction is not detected in certain other tryptophan-containing proteins. We discuss the possible implications for protein damage by this and other radical-generating reagents.
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