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Probing the interaction induced conformation transitions in acid phosphatase with cobalt ferrite nanoparticles: Relation to inhibition and bio-activity of Chlorella vulgaris acid phosphatase

Journal 2016 · IF 3.902 · 发表于 2016 年 5 月 10 日
作者: Farooq Ahmad,Xing Zhou,Hongzhou Yao,Ying Zhou,Chao Xu
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摘要

The present study explored the interaction and kinetics of cobalt ferrite nanoparticles (NPs) with acid phosphatase (ACP) by utilizing diverse range of spectroscopic techniques. The results corroborate, the CoFe 2 O 4 NPs cause fluorescence quenching in ACP by static quenching mechanism. The negative values of van’t Hoff thermodynamic expressions (Δ H   =   −0.3293   J   mol −1 K −1 and Δ G   =   −3.960   kJ   mol −1 K −1 ) corroborate the spontaneity and exothermic nature of static quenching. The positive value of ΔS (13.2893   J   mol −1 K −1 ) corroborate that major contributors of higher and stronger binding affinity among CoFe 2 O 4 NPs with ACP were electrostatic. In addition, FTIR, UV-CD, UV–vis spectroscopy and three dimensional fluorescence (3D) techniques confirmed that CoFe 2 O 4 NPs binding induces microenvironment perturbations leading to secondary and tertiary conformation changes in ACP to a great extent. Furthermore, synchronous fluorescence spectroscopy (SFS) affirmed the comparatively significant changes in microenvironment around tryptophan (Trp) residue by CoFe 2 O 4 NPs. The effect of CoFe 2 O 4 NPs on the activation kinetics of ACP was further examined in Chlorella vulgaris . Apparent Michaelis constant (Km) values of 0.57 and 26.5   mM with activation energy values of 0.538 and 3.428   kJ   mol −1 were determined without and with 200   μM CoFe 2 O 4 NPs. Apparent Vmax value of −7   Umml −1 corroborate that enzyme active sites were completely captured by the NPs leaving no space for the substrate. The results confirmed that CoFe 2 O 4 NPs ceased the activity by unfolding of ACP enzyme. This suggests CoFe 2 O 4 NPs perturbed the enzyme activity by transitions in conformation and hence the metabolic activity of ACP. This study provides the pavement for novel and simple approach of using sensitive biomarkers for sensing NPs in environment.

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