Crocetin, dimethylcrocetin, and safranal bind human serum albumin: stability and antioxidative properties.
Crocetin (CRT) and dimethylcrocetin (DMCRT) are derived from crocins which are found in the stigmas of saffron (Crocus sativus L.), while safranal is the main component of saffron's essential oil. The aim of the present study was to examine their interaction with human serum albumin in aqueous solution at physiological conditions using constant protein concentration and various ligand contents. FT-IR and UV-visible spectroscopic methods were used to determine the ligands' binding mode, the binding constant, and the effects of ligand complexation on protein secondary structure. Structural analysis showed that crocetin, dimethylcrocetin, and safranal bind nonspecifically (H-bonding) via protein polar groups with binding constants of Kcrt =2.05 (+/-0.30) x 103 M-1, Kdmcrt = 9.60 (+/-0.35) x 104 M-1, and Ksaf = 2.11 (+/-0.35) x 103 M-1. The protein secondary structure showed no major alterations at low ligand concentrations (1 microM), whereas at higher content (1 mM), decrease of alpha-helix from 55% (free HSA) to 43-45% and increase of beta-sheet from 17% (free HSA) to 18-22% and random coil 7% (free HSA) to 10-14% occurred in the ligand-HSA complexes. The results point to a partial unfolding of protein secondary structure at high ligand content. The antioxidant activity of CRT, DMCRT, and safranal was also tested by the DPPH* antioxidant activity assay, and their IC50 values were compared to that of well-known antioxidants such as Trolox and butylated hydroxy toluene (BHT). The IC50 values of CRT and safranal were 17.8 +/- 1 microg/mL and 95 +/- 1 microg/mL, respectively, while the inhibition of DMCRT reached a point of 38.8%, which corresponds to a concentration of 40 microg/mL, and then started to decrease. The IC50 values of Trolox and BHT were 5.2 +/- 1 microg/mL and 5.3 +/- 1 microg/mL, respectively.
Kanakis
Charalabos D
CD
Laboratory of Chemistry, Department of Science, Agricultural University of Athens, 75 Iera Odos, 11855 Athens, Greece.
Tarantilis
Petros A
PA
Tajmir-Riahi
Heidar Ali
HA
Polissiou
Moschos G
MG
D000975:Antioxidants / Q000494:pharmacology*; D002338:Carotenoids / Q000378:metabolism*; D053138:Cyclohexenes / Q000378:metabolism*; D004355:Drug Stability; D006801:Humans; D006860:Hydrogen Bonding; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*; D013053:Spectrophotometry; D017550:Spectroscopy, Fourier Transform Infrared; D013729:Terpenes / Q000378:metabolism*; D014801:Vitamin A / Q000031:analogs & derivatives
D000975:Antioxidants; D053138:Cyclohexenes; D012709:Serum Albumin; D013729:Terpenes; C487773:trans-sodium crocetinate; D014801:Vitamin A; D002338:Carotenoids; C087963:safranal; C519269:dimethylcrocetin