ArticleAnalytical chemistry2008
Determination of sphingosine kinase activity for cellular signaling studies.
Article in Analytical chemistry, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Thin Layer Chromatography Goes Ultrasmall to Assay Sphingosine Kinase Activation in Single Primary Leukemic Cells.Analytical chemistry · 2025Article
- "Fix and Click" for Assay of Sphingolipid Signaling in Single Primary Human Intestinal Epithelial Cells.Analytical chemistry · 2022Article
- Silicon Photomultipliers as a Low-Cost Fluorescence Detector for Capillary Electrophoresis.Analytical chemistry · 2020Article
- Application of Capillary Electrophoresis with Laser-Induced Fluorescence to Immunoassays and Enzyme Assays.Molecules (Basel, Switzerland) · 2019Review
- Sphingolipids in adipose tissue: What's tipping the scale?Advances in biological regulation · 2018Review
- Design and Application of Sensors for Chemical Cytometry.ACS chemical biology · 2018Review
- Separation of peptide fragments of a protein kinase C substrate fused to a β-hairpin by capillary electrophoresis.Analytical and bioanalytical chemistry · 2015Article
- Analysis of sphingosine kinase activity in single natural killer cells from peripheral blood.Integrative biology : quantitative biosciences from nano to macro · 2015Article
- Automated capillary electrophoresis system for fast single-cell analysis.Analytical chemistry · 2013Article
- Separation of fluorescently labeled phosphoinositides and sphingolipids by capillary electrophoresis.Journal of chromatography. B, Analytical technologies in the biomedical and life sciences · 2012Article
- A rapid assay for assessment of sphingosine kinase inhibitors and substrates.Analytical biochemistry · 2011Article
- Single-cell analysis of phosphoinositide 3-kinase and phosphatase and tensin homolog activation.Faraday discussions · 2011Article
- Microelectrophoresis platform for fast serial analysis of single cells.Electrophoresis · 2010Article
- Role of sphingosine-1-phosphate (S1P) and the S1P(2) receptor in allergen-induced, mast cell-dependent contraction of rat lung parenchymal strips.Naunyn-Schmiedeberg's archives of pharmacology · 2009Article
- Capillary electrophoresis in bioanalysis.Analytical chemistry · 2008Review
Corrections and comments
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Authors and funding
7 authors.
Funding
Abstract
Regulation of sphingosine and sphingosine-1-phosphate concentrations is of growing interest due to their importance in cellular signal transduction. Furthermore, new pharmaceutical agents moderating the intracellular and extracellular levels of sphingosine metabolites are showing promise in preclinical and clinical trials. In the present work, a quantitative assay relying on capillary electrophoresis with laser-induced fluorescence detection was developed to measure the interconversion of sphingosine and sphingosine-1-phosphate. The assay was demonstrated to be capable of determining the in vitro activity of both kinase and phosphatase using purified enzymes. The KM of sphingosine kinase for its fluorescently labeled substrate was 38 +/- 18 microM with a Vmax of 0.4 +/- 0.2 microM/min and a kcat of 3900 s-1. Pharmacologic inhibition of sphingosine kinase in a concentration-dependent manner was also demonstrated. Moreover, the fluorescent substrate was shown to be readily taken up by mammalian cells making it possible to study the endogenous activity of sphingosine kinase activity in living cells. The method was readily adaptable to the use of either bulk cell lysates or very small numbers of intact cells. This new methodology provides enhancements over standard methods in sensitivity, quantification, and manpower for both in vitro and cell-based assays.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.