Cholesterol esterase action on human high density lipoproteins and inhibition studies: detection by MALDI-TOF MS

Zschörnig, Olaf; Pietsch, Markus; Süß, Rosemarie; Schiller, Jürgen; Gütschow, Michael

doi:10.1194/jlr.m400265-jlr200

Cited by 18 publications

(13 citation statements)

References 35 publications

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“…1b) for 1 h at 37°C after the proteins were separated, electrotransferred to the membrane, and stained. Since it has been reported that the peaks of phosphatidylcholine and lysophosphatidylcholine are examined by MALDI-TOF MS, the peaks obtained in the present study are compared with evidences in the report [9]. The main peak (m/z=761) corresponds to phosphatidylcholine (C 16:0 /C 18:1 ) in Fig.…”

Section: Resultsmentioning

confidence: 81%

Reversible Inhibition of Esterase Activity After Separation and Immobilization

Sakikawa

Shimazaki

2011

Appl Biochem Biotechnol

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An inhibitor, 9-amino-1,2,3,4-tetra hydroacridine (tacrine), is a reversible inhibitor of esterases. The reversible inhibition of the enzyme activity is thought to be examined after separation and immobilization of the enzyme under non-denaturing conditions. Hydrolytic changes of phosphatidylcholine by carboxylesterase were obtained using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after the esterase was separated by non-denaturing two-dimensional electrophoresis, was immobilized to membranes and was stained by Ponceau S. The changes were inhibited after the enzyme on the membrane was treated by tacrine. Furthermore, the hydrolytic activity of the esterase was recovered after the inhibitor was washed with aspartic acid solution. These results indicate that the phosphatidylcholine hydrolysis activity of the isolated and immobilized enzyme is reversibly inhibited under non-denaturing conditions. Furthermore, this method can be developed to the production of an enzyme reactor able to regulate amounts of lipids.

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Section: Resultsmentioning

confidence: 81%

Reversible Inhibition of Esterase Activity After Separation and Immobilization

Sakikawa

Shimazaki

2011

Appl Biochem Biotechnol

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“…3a. It has been reported that the same MS peaks at m/z 496 and 524 [M + H + ] of lysophosphatidylcholine were obtained when pure phosphatidylcholine was digested by cholesterol esterase [6]. So, these peaks could represent lysophosphatidylcholine digested by phosholipase C␣ in the non-denaturing 2DE gel.…”

Section: Activity Analysis and Identification Of Phospholipase Cα Aftmentioning

confidence: 71%

“…So, there is no information whether the separated enzymes catalyze hydrolysis of the native molecules such as retinyl ester and phosphlipids. Since it has reported that enzymatic hydrolysis of lipids such as phosphatidylcholine is investigated using matrixassisted laser desorption ionisation-time of flight-mass spectrometry (MALDI-TOF-MS) [6], hydrolytic activity with phosphatidylcholine and retinyl palmitate can be investigated using MALDI-TOF-MS after cytosolic enzymes from the retina are separated using non-denaturing 2DE.…”

Section: Introductionmentioning

confidence: 99%

Analysis of hydrolytic activity of phospholipase Cα from porcine retina on retinyl ester and phosphatidylcholine using non-denaturing two-dimensional electrophoresis and mass spectrometry

Shimazaki

Kishi

Mori

et al. 2006

Journal of Chromatography B

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“…Therefore, enzymes on a membrane might catalyze the conversion of macromolecular substrates such as proteins after separation by non-denaturing 2DE. Recently, it has been reported that esterases (or lipases) hydrolyzes lipids in lipoproteins such as high density lipoprotein (HDL) and low density lipoprotein (LDL) (Hui and Howles, 2002;Zschörnig et al, 2005). As well, we previously reported the examination of carboxylesterase activity within nondenaturing 2DE after extraction from mouse liver with detergents and isolation by 2DE (Shimazaki et al, 2005).…”

Section: Introductionmentioning

confidence: 93%

Analysis of lipid hydrolytic activity by esterase on blotting membrane followed by separation using non‐denaturing two‐dimensional gel electrophoresis

Shimazaki

Miyamoto

2007

Biotech & Bioengineering

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After separation by microscale non-denaturing two-dimensional gel electrophoresis (2DE) and transferring to a blotting membrane, major proteins are detected by a staining of direct blue 71 in a neutral solution. The carboxylesterase on the membrane hydrolyzes phosphatidylcholine after the spot of carboxylesterase is excised from the membrane, and incubated with phosphatidylcholine. Lipids of human serum proteins and the purified human high density lipoprotein (HDL) are removed by enzymatic hydrolysis when human serum proteins and the purified HDL are respectively incubated with the spot of carboxylesterase on the membrane. These results indicate that carboxylesterase on the membrane hydrolyzes not only lipids such as phosphatidylcholine but also lipids of lipoproteins such as HDL after separation by the 2DE, transferring to the membrane and staining without impairing the activity. These results also indicate that a micro-immobilized enzyme reactor on the membrane can be produced when biological enzymes are separated by microscale 2DE, transferred to the membrane and stained without impairing their activities.

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Cholesterol esterase action on human high density lipoproteins and inhibition studies: detection by MALDI-TOF MS

Cited by 18 publications

References 35 publications

Reversible Inhibition of Esterase Activity After Separation and Immobilization

Reversible Inhibition of Esterase Activity After Separation and Immobilization

Analysis of hydrolytic activity of phospholipase Cα from porcine retina on retinyl ester and phosphatidylcholine using non-denaturing two-dimensional electrophoresis and mass spectrometry

Analysis of lipid hydrolytic activity by esterase on blotting membrane followed by separation using non‐denaturing two‐dimensional gel electrophoresis

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