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Analysis of Molecular Species Profiles of Ceramide-1-phosphate and Sphingomyelin Using MALDI-TOF Mass Spectrometry

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Lipids

Abstract

Ceramide-1-phosphate (C1P) is a potential signaling molecule that modulates various cellular functions in animals. It has been known that C1P with different N-acyl lengths induce biological responses differently. However, molecular species profiles of the C1P in animal tissues have not been extensively examined yet. Here, we developed a method for determination of the molecular species of a C1P using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with Phos-tag, a phosphate capture molecule. The amounts of total C1P in skin, brain, liver, kidney and small intestine of mice were determined to be 344, 151, 198, 96 and 90 pmol/g wet weight, respectively. We found a C1P species having an α-hydroxypalmitoyl residue (h-C1P, 44 pmol/g wet weight) in mouse skin. The h-C1P was detected only in the skin, and not other tissues of mice. The same analysis was applied to sphingomyelin after conversion of sphingomyelin to C1P by Streptomyces chromofuscus phospholipase D. We found that molecular species profiles of sphingomyelin in skin, kidney and small intestine of mice were similar to those of C1P in corresponding tissues. In contrast, molecular species profiles of sphingomyelin in liver and brain were quite different from those of C1P in these tissues, indicating selective synthesis or degradation of C1P in these tissues. The method described here will be useful for detection of changes in molecular species profiles of C1P and sphingomyelin.

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Abbreviations

C1P:

Ceramide-1-phosphate

h-C1P:

C1P with 2-hydroxyl fatty acyl residue

LCB:

Long-chain base

lysoPtdOH:

Lysophosphatidic acid

MALDI-TOF MS:

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

PC1P:

Phytoceramide-1-phosphate

PLD:

Phospholipase D

PtdCho:

Phosphatidylcholine

S1P:

Sphingosine-1-phosphate

THAP:

2,4,6-Trihydroxyacetophenone

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Acknowledgments

This work was partly supported by grants-in-aid from the Ministry of Education, Science, Sports, and Culture of Japan (24580185), and the research program for development of an intelligent Tokushima artificial exosome (iTEX) from Tokushima University.

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Authors and Affiliations

  1. Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima, 770-8505, Japan

    Ryouhei Yamashita, Yumika Tabata, Erina Iga, Michiyasu Nakao, Shigeki Sano, Kentaro Kogure, Akira Tokumura & Tamotsu Tanaka

  2. Department of Pharmaceutical Life Sciences, Faculty of Pharmacy, Yasuda Women’s University, Hiroshima, 731-0351, Japan

    Akira Tokumura

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  1. Ryouhei Yamashita
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  2. Yumika Tabata
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Correspondence to Tamotsu Tanaka.

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Yamashita, R., Tabata, Y., Iga, E. et al. Analysis of Molecular Species Profiles of Ceramide-1-phosphate and Sphingomyelin Using MALDI-TOF Mass Spectrometry. Lipids 51, 263–270 (2016). https://doi.org/10.1007/s11745-015-4082-0

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  • DOI: https://doi.org/10.1007/s11745-015-4082-0

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