Purification and studies on characteristics of cholinesterases from Daphnia magna

Yang, Yanxia; Niu, Lizhi; Li, Shaonan

doi:10.1631/jzus.b1200113

Cited by 11 publications

(9 citation statements)

References 56 publications

(54 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This interference means that the toxicants are not binding or inhibiting the ChE, but instead bind to other protein through various interactions. Previous studies also reported purified ChE from various samples to study the inhibition characteristics of toxicant (Li and Han, 2002;Hsiao et al, 2004;Yang et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

Purification and Anticholinesterase Sensitivity of Cholinesterase Extracted from Liver Tissue of Puntius Javanicus

Sabullah¹,

Shukor²,

Shamaan³

et al. 2015

IJAB

View full text Add to dashboard Cite

The purification of a soluble cholinesterase (ChE) from Puntius javanicus liver using affinity chromatography was studied. Affinity matrix was synthesised through the cooling system of ligands procainamide to epoxy-activated Sephacryl 6B and purification process was performed using calibrated flow rate at 0.2 mL/min. Non-denaturing electrophoresis condition was employed and the single band native form of ChE was detected at 66.267 kDa after being stained with commasie brilliant blue. ChE detection was performed using gel filtration; ZORBAX column attached to the HPLC with the flow rate of 1 mL/min. Only a single peak was detected at the retention time of 3.720. From the assay evaluation, the final purified ChE procedure displayed the highest sensitivity of detecting the anticholinesterase namely mercury, copper, malaoxon and carbofuran compared to the impure ChE and the results were further discussed in detail to the potential application of ChE from P. javanicus as a biomarker for those toxicants.

show abstract

Section: Discussionmentioning

confidence: 99%

Purification and Anticholinesterase Sensitivity of Cholinesterase Extracted from Liver Tissue of Puntius Javanicus

Sabullah¹,

Shukor²,

Shamaan³

et al. 2015

IJAB

View full text Add to dashboard Cite

show abstract

“…It does not penetrate to the active site of AChE and it inhibits BChE only. It is typically used as a reagent for rapid distinction between AChE and BChE activity in biological samples [ 51 , 52 ].…”

Section: Division Of Inhibitorsmentioning

confidence: 99%

Inhibitors of Acetylcholinesterase and Butyrylcholinesterase Meet Immunity

Pohanka

2014

IJMS

196

112

View full text Add to dashboard Cite

Acetylcholinesterase (AChE) inhibitors are widely used for the symptomatic treatment of Alzheimer’s disease and other dementias. More recent use is for myasthenia gravis. Many of these inhibitors interact with the second known cholinesterase, butyrylcholinesterase (BChE). Further, evidence shows that acetylcholine plays a role in suppression of cytokine release through a “cholinergic anti-inflammatory pathway” which raises questions about the role of these inhibitors in the immune system. This review covers research and discussion of the role of the inhibitors in modulating the immune response using as examples the commonly available drugs, donepezil, galantamine, huperzine, neostigmine and pyridostigmine. Major attention is given to the cholinergic anti-inflammatory pathway, a well-described link between the central nervous system and terminal effector cells in the immune system.

show abstract

“…The protocol was in accord with that described by Liu et al (2012a) and Yang et al (2013). Acetylthiocholine iodide and propionylthiocholine iodide were used as substrates in this protocol.…”

Section: Che Activity Measurementmentioning

confidence: 99%

“…The ChE activity of the crude enzymes was determined using propionylthiocholine iodide as substrate by the method described by Liu et al (2012a) and Yang et al (2013).…”

Section: Determination Of Cross-reactivitymentioning

confidence: 99%

See 1 more Smart Citation

Developing antibodies from cholinesterase derived from prokaryotic expression and testing their feasibility for detecting immunogen content in Daphnia magna

Liu

Yuan

2016

J. Zhejiang Univ. Sci. B

Self Cite

View full text Add to dashboard Cite

Abstract:To yield cholinesterase (ChE) from prokaryotic expression, the ChE gene that belongs to Daphnia magna was amplified by reverse transcription-polymerase chain reaction (RT-PCR) using forward primer 5'-CCCYGGNGCSAT GATGTG-3' and reverse primer 5'-GYAAGTTRGCCCAATATCT-3'. To express the gene, one sequence of the amplified DNA, which was able to encode a putative protein containing two conserved carboxylesterase domains, was connected to the prokaryotic expression vector PET-29a(+). The recombinant vector was transformed into Escherichia coil BL21 (DE3). Protein expression was induced by isopropy-D-thiogalactoside. The expressed ChE was used as an immunogen to immunize BALB/c mice. The obtained antibodies were tested for their specificity towards crude enzymes from species such as Alona milleri, Macrobrachium nipponense, Bombyx mori, Chironomus kiiensis, Apis mellifera, Eisenia foetida, Brachydanio rerio, and Xenopus laevis. Results indicated that the antibodies had specificity suitable for detecting ChE in Daphnia magna. A type of indirect and non-competitive enzyme-linked immunosorbent assay (IN-ELISA) was used to test the immunoreactive content of ChE (ChE-IR) in Daphina magna. The detection limit of the IN-ELISA was found to be 14.5 ng/ml at an antiserum dilution of 1:22 000. Results from tests on Daphnia magna exposed to sublethal concentrations of triazophos indicated a maximal induction of 57.2% in terms of ChE-IR on the second day after the animals were exposed to a concentration of 2.10 μg/L triazophos. Testing on animals acclimatized to a temperature of 16 °C indicated that ChE-IR was induced by 16.9% compared with the ChE-IR content detected at 21 °C, and the rate of induction was 25.6% at 10 °C. The IN-ELISA was also used to test the stability of ChE-IR in collected samples. Repeated freezing and thawing had no influence on the outcome of the test. All these results suggest that the polyclonal antibodies developed against the recombinant ChE are as efficient as those developed against the native ChE in detecting ChE content in Daphnia magna.

show abstract

Purification and studies on characteristics of cholinesterases from Daphnia magna

Cited by 11 publications

References 56 publications

Purification and Anticholinesterase Sensitivity of Cholinesterase Extracted from Liver Tissue of Puntius Javanicus

Purification and Anticholinesterase Sensitivity of Cholinesterase Extracted from Liver Tissue of Puntius Javanicus

Inhibitors of Acetylcholinesterase and Butyrylcholinesterase Meet Immunity

Developing antibodies from cholinesterase derived from prokaryotic expression and testing their feasibility for detecting immunogen content in Daphnia magna

Contact Info

Product

Resources

About