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Characterization of Acid Sphingomyelinase Activity in Human Cerebrospinal Fluid

BACKGROUND: As a key enzyme in sphingolipid metabolism, acid sphingomyelinase (ASM) is involved in the regulation of cell fate and signaling via hydrolysis of sphingomyelin to form ceramide. While increased activity of the lysosomal form has been associated with various pathological conditions, ther...

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Autores principales: Mühle, Christiane, Huttner, Hagen B., Walter, Silke, Reichel, Martin, Canneva, Fabio, Lewczuk, Piotr, Gulbins, Erich, Kornhuber, Johannes
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3642176/
https://www.ncbi.nlm.nih.gov/pubmed/23658784
http://dx.doi.org/10.1371/journal.pone.0062912
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author Mühle, Christiane
Huttner, Hagen B.
Walter, Silke
Reichel, Martin
Canneva, Fabio
Lewczuk, Piotr
Gulbins, Erich
Kornhuber, Johannes
author_facet Mühle, Christiane
Huttner, Hagen B.
Walter, Silke
Reichel, Martin
Canneva, Fabio
Lewczuk, Piotr
Gulbins, Erich
Kornhuber, Johannes
author_sort Mühle, Christiane
collection PubMed
description BACKGROUND: As a key enzyme in sphingolipid metabolism, acid sphingomyelinase (ASM) is involved in the regulation of cell fate and signaling via hydrolysis of sphingomyelin to form ceramide. While increased activity of the lysosomal form has been associated with various pathological conditions, there are few studies on secretory ASM limited only to cell models, plasma or serum. METHODS: An optimized assay based on a fluorescent substrate was applied to measure the ASM activity in cerebrospinal fluid (CSF) collected from mice and from 42 patients who were classified as controls based on normal routine CSF values. RESULTS: We have detected ASM activity in human CSF, established a sensitive quantitative assay and characterized the enzyme’s properties. The enzyme resembles plasmatic ASM including protein stability and Zn(2+)-dependence but the assays differ considerably in the optimal detergent concentration. Significantly increased activities in the CSF of ASM transgenic mice and undetectable levels in ASM knock-out mice prove that the measured ASM activity originates from the ASM-encoding gene SMPD1. CSF localized ASM activities were comparable to corresponding serum ASM levels at their respective optimal reaction conditions, but no correlation was observed. The large variance in ASM activity was independent of sex, age or analyzed routine CSF parameters. CONCLUSIONS: Human and mouse CSF contain detectable levels of secretory ASM, which are unrelated to serum ASM activities. Further investigations in humans and in animal models will help to elucidate the role of this enzyme in human disease and to assess its value as a potential biomarker for disease type, severity, progress or therapeutic success.
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spelling pubmed-36421762013-05-08 Characterization of Acid Sphingomyelinase Activity in Human Cerebrospinal Fluid Mühle, Christiane Huttner, Hagen B. Walter, Silke Reichel, Martin Canneva, Fabio Lewczuk, Piotr Gulbins, Erich Kornhuber, Johannes PLoS One Research Article BACKGROUND: As a key enzyme in sphingolipid metabolism, acid sphingomyelinase (ASM) is involved in the regulation of cell fate and signaling via hydrolysis of sphingomyelin to form ceramide. While increased activity of the lysosomal form has been associated with various pathological conditions, there are few studies on secretory ASM limited only to cell models, plasma or serum. METHODS: An optimized assay based on a fluorescent substrate was applied to measure the ASM activity in cerebrospinal fluid (CSF) collected from mice and from 42 patients who were classified as controls based on normal routine CSF values. RESULTS: We have detected ASM activity in human CSF, established a sensitive quantitative assay and characterized the enzyme’s properties. The enzyme resembles plasmatic ASM including protein stability and Zn(2+)-dependence but the assays differ considerably in the optimal detergent concentration. Significantly increased activities in the CSF of ASM transgenic mice and undetectable levels in ASM knock-out mice prove that the measured ASM activity originates from the ASM-encoding gene SMPD1. CSF localized ASM activities were comparable to corresponding serum ASM levels at their respective optimal reaction conditions, but no correlation was observed. The large variance in ASM activity was independent of sex, age or analyzed routine CSF parameters. CONCLUSIONS: Human and mouse CSF contain detectable levels of secretory ASM, which are unrelated to serum ASM activities. Further investigations in humans and in animal models will help to elucidate the role of this enzyme in human disease and to assess its value as a potential biomarker for disease type, severity, progress or therapeutic success. Public Library of Science 2013-05-02 /pmc/articles/PMC3642176/ /pubmed/23658784 http://dx.doi.org/10.1371/journal.pone.0062912 Text en © 2013 Mühle et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Mühle, Christiane
Huttner, Hagen B.
Walter, Silke
Reichel, Martin
Canneva, Fabio
Lewczuk, Piotr
Gulbins, Erich
Kornhuber, Johannes
Characterization of Acid Sphingomyelinase Activity in Human Cerebrospinal Fluid
title Characterization of Acid Sphingomyelinase Activity in Human Cerebrospinal Fluid
title_full Characterization of Acid Sphingomyelinase Activity in Human Cerebrospinal Fluid
title_fullStr Characterization of Acid Sphingomyelinase Activity in Human Cerebrospinal Fluid
title_full_unstemmed Characterization of Acid Sphingomyelinase Activity in Human Cerebrospinal Fluid
title_short Characterization of Acid Sphingomyelinase Activity in Human Cerebrospinal Fluid
title_sort characterization of acid sphingomyelinase activity in human cerebrospinal fluid
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3642176/
https://www.ncbi.nlm.nih.gov/pubmed/23658784
http://dx.doi.org/10.1371/journal.pone.0062912
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