Ceramide kinase regulates the production of tumor necrosis factor α (TNFα) via inhibition of TNFα-converting enzyme

Nadia F. Lamour, Dayanjan S. Wijesinghe, Jennifer A. Mietla, Katherine E. Ward, Robert V. Stahelin, Charles E. Chalfant

Research output: Contribution to journalArticle

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Abstract

Tumor necrosis factor α (TNFα) is a well known cytokine involved in systemic and acute inflammation. In this study, we demonstrate that ceramide 1-phosphate (C1P) produced by ceramide kinase (CERK) is a negative regulator of LPS-induced TNFα secretion. Specifically, bone marrow-derived macrophages isolated from CERK knock-out mice (CERK -/-) generated higher levels of TNFα than the wild-type mice (CERK +/+) in response to LPS. An increase in basal TNFα secretion was also observed in CERK -/- murine embryonic fibroblasts, which was rescued by re-expression of wild-type CERK. This effect was due to increased secretion and not transcription. The secretion of TNFα is regulated by TNFα-converting enzyme (TACE also known as ADAM17), and importantly, the activity of TACE was higher in cell extracts from CERK -/- as compared with wild type. In vitro analysis also demonstrated that C1P is a potent inhibitor of this enzyme, in stark contrast to ceramide and sphingosine 1-phosphate. Furthermore, TACE specifically bound C1P with high affinity. Finally, several putative C1P-binding sites were identified via homology throughout the protein sequence of TACE. These results indicate that C1P produced by CERK has a negative effect on the processing/secretion of TNFα via modulation of TACE activity.

Original languageEnglish (US)
Pages (from-to)42808-42817
Number of pages10
JournalJournal of Biological Chemistry
Volume286
Issue number50
DOIs
StatePublished - Dec 16 2011

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Tumor Necrosis Factor-alpha
Enzymes
Amino Acid Sequence Homology
Macrophages
Ceramides
Enzyme Inhibitors
Transcription
Fibroblasts
ceramide kinase
Cell Extracts
Knockout Mice
ceramide 1-phosphate
Bone
Binding Sites
Modulation
Cytokines
Inflammation
Processing
Proteins
ADAM17 Protein

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Ceramide kinase regulates the production of tumor necrosis factor α (TNFα) via inhibition of TNFα-converting enzyme. / Lamour, Nadia F.; Wijesinghe, Dayanjan S.; Mietla, Jennifer A.; Ward, Katherine E.; Stahelin, Robert V.; Chalfant, Charles E.

In: Journal of Biological Chemistry, Vol. 286, No. 50, 16.12.2011, p. 42808-42817.

Research output: Contribution to journalArticle

Lamour, Nadia F. ; Wijesinghe, Dayanjan S. ; Mietla, Jennifer A. ; Ward, Katherine E. ; Stahelin, Robert V. ; Chalfant, Charles E. / Ceramide kinase regulates the production of tumor necrosis factor α (TNFα) via inhibition of TNFα-converting enzyme. In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 50. pp. 42808-42817.
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