33 research outputs found
Hydrolysis improves the inhibition efficacy of bovine lactoferrin against infection by SARS-CoV-2 pseudovirus
No full textThe entry of SARS-CoV-2 into host cells may involve the spike protein cleavage by cathepsin L (CTSL). Certain food proteins such as lactoferrin (Lf) inhibit CTSL. The current study investigated the impact of hydrolysis (0–180 min) by proteinase K on electrophoretic pattern, secondary structure, cathepsin inhibitory and SARS-CoV-2 pseudovirus infectivity inhibitory of bovine Lf. Gel electrophoresis indicated that hydrolysis cut Lf molecules to half lobes (∼40 kDa) and produced peptides ≤18 kDa. Approximation of the secondary structural features through analysis of the second-derivative amide I band collected by infra-red spectroscopy suggested a correlative–causative relationship between cathepsin inhibition and the content of helix-unordered structures in Lf hydrolysate. The half maximal inhibitory concentration (IC(50)) of Lf hydrolysed for 90 min (H90) against CTSL was about 100 times smaller than that of the Lf hydrolysed for 0 min (H0). H90 had also double activity against SARS-CoV-2 pseudo-types infectivity compared with H0
Binding sensorgram for tannic acid interaction with immobilized AChE.
No full textIncreasing concentrations of tannic acid from 0.78 μM-12.5 μM were injected over the enzyme surface at 25°C. The flow rate is maintained at 45 μl min-1. Contact time and dissociation time was kept at 120 s and 200 s. Regeneration was carried out using 10 mM glycine HCl with pH 3 for 30 s and at 30 μl min-1. The data analysis was done using Biacore X100 evaluation software ver 2.0.2 and data was fit to two state. The resulting equilibrium dissociation constants KD, kinetic association kon and dissociation koff rates are given in Table 2.</p
Binding sensorgram for quercetin interaction with immobilized AChE.
No full textIncreasing concentrations of quercetin from 50 μM-800 μM were injected over the enzyme surface at 25°C. The flow rate is maintained at 45 μl min-1. Contact time and dissociation time was kept at 120 s and 200 s. Regeneration was carried out using 10 mM glycine HCl with pH 3 for 30 s and at 30 μl min-1. The data analysis was done using Biacore X100 evaluation software ver 2.0.2 and data was fit to two state. The resulting equilibrium dissociation constants KD, kinetic association kon and dissociation koff rates are given in Table 2.</p
Binding sensorgram for galamer4 interaction with immobilized AChE.
No full textIncreasing concentrations of galanthamine tablet from 50 μM-800 μM were injected over the enzyme surface at 25°C. Flow rate is maintained at 45 μl min-1. Contact time and dissociation time was kept at 120 s and 200 s. Regeneration was carried out using 10 mM glycine HCl with pH 3 for 30 s and at 30 μl min-1. The data analysis was done using Biacore X100 evaluation software ver 2.0.2 and data was fit to two state. The resulting equilibrium dissociation constants KD, kinetic association kon and dissociation koff rates are given in Table 2.</p
Binding sensorgram for tannic acid and galanthamine hydrobromide interaction with immobilized AChE.
No full textIncreasing concentrations of tannic acid from 3.12 μM-50 μM and galanthamine hydrobromide from 50 μM-800 μM were injected over the enzyme surface at 25°C. The flow rate is maintained at 45 μl min-1. Contact time and dissociation time was kept at 120 s and 200 s. Regeneration was carried out using 10 mM glycine HCl with pH 3 for 30 s and at 30 μl min-1. The data analysis was done using Biacore X100 evaluation software ver 2.0.2 and data was fit to heterogeneous analyte fit. The resulting equilibrium dissociation constants are given in Table 3.</p
Comparative biophysical characterization: A screening tool for acetylcholinesterase inhibitors.
No full textAmong neurodegenerative diseases, Alzheimer's disease (AD) is one of the most grievous disease. The oldest cholinergic hypothesis is used to elevate the level of cognitive impairment and acetylcholinesterase (AChE) comprises the major targeted enzyme in AD. Thus, acetylcholinesterase inhibitors (AChEI) constitutes the essential remedy for the treatment of AD. The study aims to evaluate the interactions between natural molecules and AChE by Surface Plasmon Resonance (SPR). The molecules like alkaloids, polyphenols and substrates of AChE have been considered for the study with a major emphasis on affinity and kinetics. To better understand the activity of small molecules, the investigation is supported by both experimental and theoretical approach such as fluorescence, Circular Dichroism (CD) and molecular docking studies. Amongst the screened ones tannic acid showed promising results compared with others. The methodology followed here have highlighted many molecules with a higher affinity towards AChE and these findings may take lead molecules generated in preclinical studies to treat neurodegenerative diseases. Additionally, we suggest a unique signature for the heterogeneous analyte model using competitive experiments for analyzing simultanous interactions of both the analytes
Comparative biophysical characterization: A screening tool for acetylcholinesterase inhibitors - Fig 24
No full textIn silico docking of crystal structure of human recombinant AChE (4EY7) with different molecules k) ATCI and l) carbachol were used in these docking studies. All the molecules were taken from Zinc database. UCSF Chimera 1.12 software was used for visualization of the results and creating images.</p
Comparative biophysical characterization: A screening tool for acetylcholinesterase inhibitors - Fig 2
No full texta) Effect of molecules- quercetin, rutin hydrate, lycorine hydrochloride, caffeine, carbachol, pyrogallol and crocin concentrations on the AChE activity using ATCI as substrate. The reaction mixture contained 7.5 mM of the substrate in a final concentration of 200 mM phosphate buffer saline (PBS), pH 7.7 at 37°C. b) Effect of molecules- tannic acid, catechin hydrate, galanthamine hydrobromide and galanthamine tablet concentrations on the AChE activity using ATCI as substrate. The reaction mixture contained 7.5 mM of the substrate in a final concentration of 200 mM phosphate buffer saline (PBS), pH 7.7 at 37°C.</p
IC<sub>50</sub> values of different small molecules on AChE.
No full textIC50 values of different small molecules on AChE.</p
Binding sensorgram for lycorine hydrochloride interaction with immobilized AChE.
No full textIncreasing concentrations of lycorine hydrochloride 50 μM-800 μM from were injected over enzyme surface. The flow rate is maintained at 45 μl min-1. Contact time and dissociation time was kept at 120 s and 200 s. Regeneration was carried out using 10 mM glycine HCl with pH 3 for 30 s and at 30 μl min-1. The data analysis was done using Biacore X100 evaluation software ver 2.0.2 and data was fit to two state. The resulting equilibrium dissociation constants KD, kinetic association kon and dissociation koff rates are given in Table 2.</p
