Interaction of 7-hydroxyflavone with human serum albumin: a spectroscopic study.
Numerous recent investigations have revealed that various synthetic as well as therapeutically active natural flavonoids possess novel luminescence properties that can serve as highly sensitive monitors for exploring their interactions with relevant physiological targets. Here we report a detailed study on the interactions of the model flavone, 7-hydroxyflavone (7HF) with the plasma protein human serum albumin (HSA), employing electronic absorption, fluorescence (steady state and time resolved) and induced circular dichroism (ICD) spectroscopy. The spectral data indicate that in the protein matrix, the neutral 7HF molecules are predominantly transformed to a conjugate anion (7HFA) by a proton abstraction in the ground state. The protein (HSA) environment induces dramatic enhancements in the fluorescence emission intensity, anisotropy (r) and lifetime (tau) values, as well as pronounced changes in the fluorescence excitation and emission profiles of the fluorophore. Moreover, evidence for efficient Förster type resonance energy transfer (FRET, from tryptophan to 7HFA) is presented, from which we infer that the binding site of 7HF in HSA is proximal (estimated distance, R=23.6A) to the unique tryptophan - 214 residue present in the inter-domain (between IIA and IIIA domains) loop region of the protein. The binding constant (K=9.44x10(4)M(-1)) and the Gibbs free energy change (DeltaG=-28.33kJ/mol) for 7HFA-HSA interaction have been estimated from the emission data. Finally, the near-UV circular dichroism (CD) studies show that the electronic transitions of 7HF are strongly perturbed on binding to the chiral host (HSA), leading to the appearance of ICD bands. Implications of these results are discussed.
Banerjee
Anwesha
A
Biophysics Division, Saha Institute of Nuclear Physics 1/AF, Bidhannagar, Kolkata 700 064, India.
Basu
Kaushik
K
Sengupta
Pradeep K
PK
D002942:Circular Dichroism; D005419:Flavonoids / Q000737:chemistry*; D006801:Humans; D015394:Molecular Structure; D012709:Serum Albumin / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics; D013997:Time Factors
D005419:Flavonoids; D012709:Serum Albumin; C429845:7-hydroxyflavone