In the present investigation, an attempt has been made to study the interaction of newly synthesized bioactive compound 3-pyrazolyl 2-pyrazoline (PZ) with model transport proteins, bovine serum albumin (BSA), and human serum albumin (HSA) employing steady state and time-resolved fluorescence technique. We have focused on fluorescence resonance energy transfer (FRET) between excited tryptophan in transport proteins to transport-proteins-bound PZ. An efficient Forster-type resonance energy transfer from the tryptophan residues to PZ indicates that PZ binds in the vicinity of the tryptophan residue. Binding of protein to that bioactive compound without changing conformation of primary and secondary structure of protein has been monitored using circular dichroism (CD) study.
In an equimolar ratio the human telomeric oligonucleotides d[AGGG(TTAGGG)(3)] and d[(CCCTAA)(3)CCCT] formed mixed structures of duplex and tetraplex in bis(2-ethylhexyl)sulfosuccinate reverse micelles; only the duplex was observed in aqueous buffer. This finding suggests that heterogeneous confined media in the cell nucleus might induce a significant fraction of the telomeric region of genomic DNA to adopt non-canonical tetraplex structure.
The photophysical behavior of 3-pyrazolyl-2-pyrazoline derivative (PZ), a newly synthesized biologically active compound has been studied in micellar solutions of anionic sodium dodecyl sulfate (SDS), cationic cetyl trimethylammonium bromide (CTAB) and nonionic p- tert-octylphenoxy polyoxyethanol (Triton X-100, TX-100) micelle using steady state and time-resolved fluorescence spectroscopy technique. Influence of the micelles on the photophysics of PZ has also been investigated using different approaches. The location of the fluorophore PZ in the micelle has been identified by cetyl pyridinium chloride (CpCl) induced fluorescence quenching and micropolarity surrounding that fluorophore in micellar solution. The effect of urea on the steady state fluorescence and relaxation dynamics of the micelle bound probe has also been observed. The results have been interpreted in terms of the model that urea displaces water molecules from the micellar interface and the consequent destabilization leads to the expulsion of the probe molecules from the interfacial region. An attempt has been made to determine probe sensing microviscosities for these micellar microenvironments in the light of average reorientation times of the probe PZ.
Four nucleic acid duplexes-DNA/RNA hybrid, RNA/DNA hybrid, RNA duplex, and DNA duplex-were studied under molecular crowding conditions of osmolytes. Destabilization of duplexes (ΔΔG°(25)) indicated that the ΔΔG°(25) values of hybrids were intermediate between those of DNA and RNA duplexes. In the presence of polyethylene glycol 200, the ΔΔG°(25) values were estimated to be +3.0, +3.5, +3.5, and +4.1 kcal mol(-1) for the DNA duplex, DNA/RNA hybrid, RNA/DNA hybrid, and RNA duplex, respectively. Differences in the number of water molecules taken up (-Δn(w)) upon duplex formations between 0 and 37 °C (Δ(-Δn(w))) were estimated to be 44.8 and 59.7 per duplex structure for the DNA/RNA and RNA/DNA hybrids, respectively. While the Δ(-Δn(w)) value for the DNA/RNA hybrid was intermediate between those of the DNA (26.1) and RNA (59.2) duplexes, the value for RNA/DNA hybrid was close to that of RNA duplex. These differences in the thermodynamic parameters and hydration are probably a consequence of the enhanced global flexibility of the RNA/DNA hybrid structure relative to the DNA/RNA hybrid structure observed in molecular dynamics simulations. This molecular crowding study provides information not only on hydration but also on the flexibility of the conformation of nucleic acid duplexes.
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