Sensitivity of bovine retinal acetylcholinesterase (E.C. 3.1.1.7) toward tacrine: Kinetic characterization

Al‐Jafari, Abdulaziz A.; Kamal, Mohammad Amjad; Alhomida, Abdullah S.

doi:10.1002/(sici)1099-0461(1998)12:4<245::aid-jbt7>3.0.co;2-l

Cited by 12 publications

(1 citation statement)

References 25 publications

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“…Having shown that the DIOS‐TOFMS method is quantitative, a series of experiments was performed to characterize the enzyme kinetics for acetylcholine esterase and the inhibitor tacrine. Using DIOS‐TOFMS, the Michaelis constant for acetylcholine esterase was found to be 98 μM, in good agreement with reported values 21,22. The 50% inhibition constant, or IC 50 , for tacrine was found to be 32.0 nM by DIOS‐TOFMS and 31.8 nM by LC/MS/MS.…”

Section: Discussionsupporting

confidence: 85%

Comparison of desorption/ionization on silicon (DIOS) time‐of‐flight and liquid chromatography/tandem mass spectrometry for assaying enzyme‐inhibition reactions

Wall

Finch

Cohen

2004

Rapid Comm Mass Spectrometry

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Desorption/ionization on silicon with time-of-flight mass spectrometry (DIOS-TOFMS) was employed for rapid quantification of small molecules and the measurement of critical constants used in enzyme kinetics studies. The approach was used to determine the Michaelis constant (Km) for acetylcholine esterase and the 50% inhibition constant (IC50) for tacrine. The Km was determined from a Lineweaver-Burk plot to be 98 microM. The IC50 was determined by assaying acetylcholine esterase activity with a range of tacrine concentrations, and measuring the amount of choline produced at a fixed time point by either DIOS-TOFMS or by liquid chromatography tandem mass spectrometry (LC/MS/MS). DIOS-TOFMS indicated an IC50 of 32.0 nM while LC/MS/MS gave a value of 31.8 nM. The excellent correlation with the reported IC50 values, ranging from 30.0 to 50.0 nM, and equivalence with the LC/MS/MS results, confirms that the DIOS-TOFMS method can be used for rapid and specific quantification of enzyme-inhibition assays.

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Section: Discussionsupporting

confidence: 85%

Comparison of desorption/ionization on silicon (DIOS) time‐of‐flight and liquid chromatography/tandem mass spectrometry for assaying enzyme‐inhibition reactions

Wall

Finch

Cohen

2004

Rapid Comm Mass Spectrometry

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Micro-Scale Extraction and Analysis of Intact Carboxylesterase after Trapping on an Immunoaffinity Membrane Surface

Kimura

Shimazaki

2014

Appl Biochem Biotechnol

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Porcine liver carboxylesterase was captured using an immunoaffinity membrane, which was prepared by separating an anti-porcine esterase antibody using non-denaturing two-dimensional electrophoresis, followed by transfer to a polyvinylidene difluoride membrane and staining. The activity of this esterase was 0.008 units after it was captured in the tiny spaces (4 mm(2)) of this membrane and eluted by rinsing with 5 μL of aspartic acid solution. The molecular mass of the eluted esterase was m/z 61,885 according to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after the purification of this enzyme from the porcine liver cytosol. The purified enzyme's activity was inhibited by 6,9-diamino-2-ethoxyacridine, and this inhibition was retained even after extracting the enzyme from the immunoaffinity membrane. These results indicate that micro-scale extraction and analysis of a carboxylesterase are possible when the enzyme is trapped using an immunoaffinity membrane and eluted with aspartic acid.

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Specific Cholinesterase Inhibitors: A Potential Tool to Assist in Management of Alzheimer Disease

Grieg

Kamal

Jabir

et al. 2014

Drug Design and Discovery in Alzheimer's Disease

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Sensitivity of bovine retinal acetylcholinesterase (E.C. 3.1.1.7) toward tacrine: Kinetic characterization

Cited by 12 publications

References 25 publications

Comparison of desorption/ionization on silicon (DIOS) time‐of‐flight and liquid chromatography/tandem mass spectrometry for assaying enzyme‐inhibition reactions

Comparison of desorption/ionization on silicon (DIOS) time‐of‐flight and liquid chromatography/tandem mass spectrometry for assaying enzyme‐inhibition reactions

Micro-Scale Extraction and Analysis of Intact Carboxylesterase after Trapping on an Immunoaffinity Membrane Surface

Specific Cholinesterase Inhibitors: A Potential Tool to Assist in Management of Alzheimer Disease

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