Fas-activated serine/threonine kinase (FASTK) is a mitochondria-associated nuclear protein that inhibits Fas- and UV-induced apoptosis. This protein is generally activated during Fas-mediated apoptosis by phosphorylating a nuclear RNA-binding protein T-cell intracellular antigen-1 and thus considered as a modulator of apoptosis. In the present study, we have examined the equilibrium unfolding and conformational stability of the kinase domain of FASTK (FASTK<sub>353-444</sub><b>)</b>. The kinase domain of FASTK<sub>353-444</sub> was cloned, expressed, and purified. The folding ↔ unfolding transitions of urea-induced denaturation was monitored with the help of circular dichroism, intrinsic fluorescence, and UV absorption spectroscopies. Analysis of transition curves obtained from different probes revealed a coincidence of denaturation curves, suggesting that folding/unfolding of FASTK follows a two-state process with the midpoint (<i>C</i><sub>m</sub>) value at 3.50 ± 0.1 M. Urea-induced denaturation curves were further analyzed to estimate change in the Gibbs free energy in the absence of urea (Δ<i>G</i><sub>D</sub><sup>0</sup>) associated with the equilibrium of denaturation. To get atomistic insights into the urea-induced denaturation of FASTK, we performed an all-atom molecular dynamics simulation for 100 ns. A close agreement was noticed between experimental and computational studies. This study will help to understand the unfolding mechanism and structural stability of the kinase domain of FASTK.Communicated by Ramaswamy H. Sarma.
Purpose – To discuss synthesis and evaluation of organo‐metallic chalcones as second‐order nonlinear optical (SONLO) materials.
Here, the voltammetric electrochemical approach was applied to detect uric acid (UA) in a conductive sensing medium (phosphate buffer solution-PBS) by using PbO-doped NiO nanocomposites (NCs)-decorated glassy carbon electrode (GCE) performing as working electrode. The wet-chemically prepared PbO-doped NiO NCs were subjected to characterization by the implementation of XRD, FESEM, XPS, and EDS analysis. The modified GCE was used to detect uric acid (UA) in an enzyme-free conductive buffer (PBS) of pH = 7.0. As the outcomes of this study reveal, it exhibited good sensitivity of 0.2315 µAµM<sup>-1</sup>cm<sup>-2</sup> and 0.2233 µAµM<sup>-1</sup>cm<sup>-2</sup>, corresponding to cyclic (CV) and differential pulse (DPV) voltammetric analysis of UA, respectively. Furthermore, the proposed UA sensor showed a wider detection (0.15~1.35 mM) range in both electrochemical analysis methods (CV & DPV). In addition, the investigated UA sensor displayed appreciable limit of detection (LOD) of 41.0 ± 2.05 µM by CV and 43.0 ± 2.14 µM by DPV. Good reproducibility performance, faster response time and long-time stability in detection of UA were perceived in both electrochemical analysis methods. Finally, successful analysis of the bio-samples was performed using the recovery method, and the results were found to be quite acceptable in terms of accuracy. Thus, the findings indicate a reliable approach for the development of 5th generation biosensors using metal-oxides as sensing substrate to fulfill the requirements of portable use for in situ detection.
In this study, poly(luminol-<i>co</i>-1,8-diaminonaphthalene) (PLim-DAN) was synthesized and subsequently modified with MWCNTs and CeO<sub>2</sub> NPs. The synthesized nanocomposites were analyzed using IR, SEM, TEM, and XRD. Furthermore, a comprehensive set of thermal behavior measurements were taken using TGA/DTG analysis. Next, the electroactivity of the developed nanocomposites was tested as an electrochemical sensor to measure the concentration of Cr<sup>3+</sup> ions in phosphate buffers. The GCE adapted with the PLim-DAN/CeO<sub>2</sub>/CNTs-10% nanocomposite (NC) exhibited the highest current response among the other compositions and copolymers. The fabricated nanocomposite sensor showed high sensitivity, with a value of 19.78 μA μM<sup>-1</sup> cm<sup>-2</sup>, and a low detection limit of 4.80 ± 0.24 pM. The analytical performance was evaluated by plotting a current calibration curve <i>versus</i> the concentration of Cr<sup>3+</sup> ions. It was found to be linear (<i>R</i><sup>2</sup> = 0.9908) over the range of 0.1 nM to 0.1 mM, identified as the linear dynamic range (LDR). This electrochemical sensor demonstrated that it could be a useful tool for environmental monitoring by accurately detecting and measuring carcinogenic Cr<sup>3+</sup> ions in real-world samples.