MilliporeSigma

Inactivation of Alpha-2-Macroglobulin by Photo-Illuminated Gallic Acid.

Journal of fluorescence (2019-07-22)
Tooba Siddiqui, Mohammad Khalid Zia, Syed Saqib Ali, Haseeb Ahsan, Fahim Halim Khan
ABSTRACT

Gallic acid is a naturally occurring plant polyphenol found in green tea and various fruits. Under certain conditions gallic acid exhibits pro-oxidant characteristics rather than its well known antioxidant property. In the present work, we explored the interaction of gallic acid with sheep alpha-2-macroglobulin (α2M) in the presence of light and determined the functional alteration and conformational modifications induced in α2M structure. α2M is a highly abundant homotetrameric antiproteinase glycoprotein having diverse functions. Our result suggests α2M loses almost 54% of its proteinase inhibitory activity after 2 h incubation with gallic acid in presence of light. The inactivation of α2M was due to photodynamic generation of superoxide radical and hydrogen peroxide by gallic acid. The UV/visible absorption spectra of α2M showed increase in absorbance due to complex formation with gallic acid. Intrinsic fluorescence study shows that α2M-gallic acid interaction leads to quenching of fluorescence intensity of α2M and the mechanism of quenching is found to be static in nature. Synchronous fluorescence measurements reveal that gallic acid interaction leads to change in the microenvironment around tryptophan residues of α2M. Moreover, Fourier transform infrared spectroscopy and circular dichroism spectra suggests perturbation in secondary structure of α2M. Binding parameters were investigated by spectroscopic as well as calorimetric measurements. Negative value of enthalpy change and Gibbs free energy confirms the binding process to be exothermic and spontaneous.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Nα-Benzoyl-DL-arginine 4-nitroanilide hydrochloride, ≥98%