A capillary electrophoresis method with dynamic pH junction stacking for the monitoring of cerebroside sulfotransferase

Li, Wenjin; Zech, Isabell; Gieselmann, Volkmar; Müller, Christian

doi:10.1016/j.chroma.2015.06.053

Cited by 7 publications

(12 citation statements)

References 39 publications

(49 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In conclusion, we synthesised and investigated selected analogues of the natural CST substrate galactosylceramide (3), which were related to the artificial substrate KRN7000 (18). Our aim was to study structure-activity relationships for substrates of the enzyme, and to identify competitive inhibitors.…”

Section: Discussionmentioning

confidence: 99%

“…The human CST enzyme was obtained by heterologous expression in CHO cells in analogy to a described procedure 19 . The CE assay method has previously been reported 18 .…”

Section: Biological Evaluationmentioning

confidence: 99%

“…K i values were calculated according to the Cheng-Prusoff equation 20 . The substrate concentration was 100 mM of galactosylceramide (3), and 466 mM of KRN7000 (18), respectively, and the concentration of the cofactor PAPS was 30 lM. Substrate conversion was strictly controlled to be below 15%.…”

Section: Investigation Of Cst Inhibitorsmentioning

confidence: 99%

“…Thus, we investigated further a-galactosyl-lipids (11)(12)(13)(14)(15)(16)(17)(19)(20)(21)(22) derived from KRN7000 (18) mainly with modification of the fatty acid residue. KRN7000 (18) had previously been found to display immunostimulatory and antitumor activity in several in vivo models, and was advanced to clinical trials [29][30][31] .…”

Section: Substratesmentioning

confidence: 99%

See 3 more Smart Citations

Synthesis and structure-activity relationships of cerebroside analogues as substrates of cerebroside sulphotransferase and discovery of a competitive inhibitor

Guillaume²,

Wachsmann

et al. 2020

Journal of Enzyme Inhibition and Medicinal Chemistry

Self Cite

View full text Add to dashboard Cite

Metachromatic leukodystrophy (MLD) is a rare genetic disease characterised by a dysfunction of the enzyme arylsulphatase A leading to the lysosomal accumulation of cerebroside sulphate (sulphatide) causing subsequent demyelination in patients. The enzyme galactosylceramide (cerebroside) sulphotransferase (CST) catalyses the transfer of a sulphate group from 3 0-phosphoadenosine-5'-phosphosulphate (PAPS) to cerebrosides producing sulphatides. Substrate reduction therapy for arylsulphatase A by inhibition of CST was proposed as a promising therapeutic approach. To identify competitive CST inhibitors, we synthesised and investigated analogues of the substrate galactosylceramide with variations at the anomeric position, the acyl substituent and the carbohydrate moiety, and investigated their structure-activity relationships. While most of the compounds behaved as substrates, a-galactosylceramide 16 was identified as the first competitive CST inhibitor. Compound 16 can serve as a new lead structure for the development of drugs for the treatment of this devastating disease, MLD, for which small molecule therapeutics are currently not available.

show abstract

Section: Discussionmentioning

confidence: 99%

“…The human CST enzyme was obtained by heterologous expression in CHO cells in analogy to a described procedure 19 . The CE assay method has previously been reported 18 .…”

Section: Biological Evaluationmentioning

confidence: 99%

Section: Investigation Of Cst Inhibitorsmentioning

confidence: 99%

Section: Substratesmentioning

confidence: 99%

See 2 more Smart Citations

Synthesis and structure-activity relationships of cerebroside analogues as substrates of cerebroside sulphotransferase and discovery of a competitive inhibitor

Guillaume²,

Wachsmann

et al. 2020

Journal of Enzyme Inhibition and Medicinal Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…A method for the monitoring of cerebroside sulfotransferase was developed by Li et al. . A sample matrix consisting of 5 mM sodium phosphate buffer with 0.0001% polybrene at pH 7.4 and a BGE containing 75 mM sodium phosphate buffer at pH 5.6 with 0.002% polybrene were utilized to achieve the stacking effect.…”

Section: Stacking By Ph Junction and Similar Techniquesmentioning

confidence: 99%

Recent progress of sample stacking in capillary electrophoresis (2014–2016)

et al. 2016

View full text Add to dashboard Cite

The term "sample stacking" comprises a relatively broad spectrum of techniques that already form an almost inherent part of the methodology of CZE. Their principles are different but the effect is the same: concentration of a diluted analyte into a narrow zone and considerable increase of the method sensitivity. This review brings a survey of papers on electrophoretic sample stacking published approximately since the second quarter of 2014 till the first quarter of 2016. It is organized according to the principles of the stacking methods and includes chapters aimed at the concentration adjustment principle (Kohlrausch stacking), techniques based on pH changes, micellar methods, and other stacking techniques. Not reviewed are papers on transient ITP that are covered by another review in this issue.

show abstract

Advances in screening enzyme inhibitors by capillary electrophoresis

Wang

Yang

2019

Electrophoresis

View full text Add to dashboard Cite

Capillary electrophoresis (CE) has attracted lots of attention due to its simplicity, low sample consumption, low solvent volume, high resolution, and high speed. Based on these advantages, it has been widely used in enzyme inhibitor screening. There are two main operation modes on enzyme inhibitor screening: off‐line (precapillary enzyme assays) in which process CE was used as an analytical tool; online (in‐capillary enzyme assays) which combined the sample injection, mix, reaction, separation, and detection within a single run. Additionally, diverse of new materials were introduced to immobilize enzyme, which has been coupled with CE for the study of enzyme activity and its inhibitor screening. This review gives an overview of the developments and applications for the CE‐based enzyme inhibitor screening.

show abstract

A capillary electrophoresis method with dynamic pH junction stacking for the monitoring of cerebroside sulfotransferase

Cited by 7 publications

References 39 publications

Synthesis and structure-activity relationships of cerebroside analogues as substrates of cerebroside sulphotransferase and discovery of a competitive inhibitor

Synthesis and structure-activity relationships of cerebroside analogues as substrates of cerebroside sulphotransferase and discovery of a competitive inhibitor

Recent progress of sample stacking in capillary electrophoresis (2014–2016)

Advances in screening enzyme inhibitors by capillary electrophoresis

Contact Info

Product

Resources

About