Structures and biological activity of phosphorylated dihydroceramides of Porphyromonas gingivalis

Frank C. Nichols, Birgit Riep, Ji Young Mun, Martha D Morton, Mike T. Bojarski, Floyd E. Dewhirst, Michael B. Smith

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Porphyromanas gingivalis, a recognized periodontal pathogen, synthesizes free ceramides as well as other phosphorylated ceramide lipids. The purpose of this study was to separate complex lipids of P. gingivalis by high-performance liquid chromatography (HPLC) and determine the structures and biological activities of the major ceramide classes. Using gas chromatography-mass spectrometry, electrospray tandem mass spectrometry (ESI-MS/MS) and NMR analyses, three major classes of dihydroceramides were identified in specific HPLC fractions, with all classes containing the same dihydroceramide base structures (3-OH isoC17:10 in amide linkage to saturated long-chain bases of 17, 18, or 19 carbons). The free dihydroceramide class recovered in HPLC fractions 7-8 revealed little biological activity. HPLC fraction 20 dihydroceramides, substituted with 1-O-phosphoglycerol and isoC15:0 linked to the hydroxyl of 3-OH isoC17:0, significantly potentiated interleukin-1β (IL-1β) (-mediated prostaglandin secretion and produced marked alterations in fibroblast morphology. HPLC fraction 28 dihydroceramides, substituted with 1-O-phosphoethanolamine, demonstrated little capacity to potentiate IL-1β-mediated prostaglandin secretion. The novel phosphorylated dihydroceramides synthesized by P. gingivalis demonstrate varying biological activities based on the phosphorylated head group substitution and/or the addition of esterified fatty acid. These results also demonstrate the strong virulence capacity of phosphoglycerol dihydroceramides of P. gingivalis to promote inflammatory factor secretion from IL-1 β-treated fibroblasts and to produce marked alterations in cell morphology in culture.

Original languageEnglish (US)
Pages (from-to)2317-2330
Number of pages14
JournalJournal of Lipid Research
Volume45
Issue number12
DOIs
StatePublished - Dec 1 2004

Fingerprint

Porphyromonas gingivalis
Bioactivity
High performance liquid chromatography
High Pressure Liquid Chromatography
Ceramides
Interleukin-1
Fibroblasts
Prostaglandins
Mass spectrometry
Lipids
Pathogens
Tandem Mass Spectrometry
dihydroceramide
Amides
Gas chromatography
Hydroxyl Radical
Gas Chromatography-Mass Spectrometry
Virulence
Substitution reactions
Fatty Acids

Keywords

  • Electrospray MS/MS
  • Gas chromatography-mass spectrometry
  • Gingival fibroblast
  • Interleukin-1β
  • Long-chain base
  • Phosphoethanolamine ceramide
  • Phosphoglycerol dihydroceramide
  • Prostaglandin E

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

Cite this

Structures and biological activity of phosphorylated dihydroceramides of Porphyromonas gingivalis. / Nichols, Frank C.; Riep, Birgit; Mun, Ji Young; Morton, Martha D; Bojarski, Mike T.; Dewhirst, Floyd E.; Smith, Michael B.

In: Journal of Lipid Research, Vol. 45, No. 12, 01.12.2004, p. 2317-2330.

Research output: Contribution to journalArticle

Nichols, Frank C. ; Riep, Birgit ; Mun, Ji Young ; Morton, Martha D ; Bojarski, Mike T. ; Dewhirst, Floyd E. ; Smith, Michael B. / Structures and biological activity of phosphorylated dihydroceramides of Porphyromonas gingivalis. In: Journal of Lipid Research. 2004 ; Vol. 45, No. 12. pp. 2317-2330.
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AU - Nichols, Frank C.

AU - Riep, Birgit

AU - Mun, Ji Young

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AU - Bojarski, Mike T.

AU - Dewhirst, Floyd E.

AU - Smith, Michael B.

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N2 - Porphyromanas gingivalis, a recognized periodontal pathogen, synthesizes free ceramides as well as other phosphorylated ceramide lipids. The purpose of this study was to separate complex lipids of P. gingivalis by high-performance liquid chromatography (HPLC) and determine the structures and biological activities of the major ceramide classes. Using gas chromatography-mass spectrometry, electrospray tandem mass spectrometry (ESI-MS/MS) and NMR analyses, three major classes of dihydroceramides were identified in specific HPLC fractions, with all classes containing the same dihydroceramide base structures (3-OH isoC17:10 in amide linkage to saturated long-chain bases of 17, 18, or 19 carbons). The free dihydroceramide class recovered in HPLC fractions 7-8 revealed little biological activity. HPLC fraction 20 dihydroceramides, substituted with 1-O-phosphoglycerol and isoC15:0 linked to the hydroxyl of 3-OH isoC17:0, significantly potentiated interleukin-1β (IL-1β) (-mediated prostaglandin secretion and produced marked alterations in fibroblast morphology. HPLC fraction 28 dihydroceramides, substituted with 1-O-phosphoethanolamine, demonstrated little capacity to potentiate IL-1β-mediated prostaglandin secretion. The novel phosphorylated dihydroceramides synthesized by P. gingivalis demonstrate varying biological activities based on the phosphorylated head group substitution and/or the addition of esterified fatty acid. These results also demonstrate the strong virulence capacity of phosphoglycerol dihydroceramides of P. gingivalis to promote inflammatory factor secretion from IL-1 β-treated fibroblasts and to produce marked alterations in cell morphology in culture.

AB - Porphyromanas gingivalis, a recognized periodontal pathogen, synthesizes free ceramides as well as other phosphorylated ceramide lipids. The purpose of this study was to separate complex lipids of P. gingivalis by high-performance liquid chromatography (HPLC) and determine the structures and biological activities of the major ceramide classes. Using gas chromatography-mass spectrometry, electrospray tandem mass spectrometry (ESI-MS/MS) and NMR analyses, three major classes of dihydroceramides were identified in specific HPLC fractions, with all classes containing the same dihydroceramide base structures (3-OH isoC17:10 in amide linkage to saturated long-chain bases of 17, 18, or 19 carbons). The free dihydroceramide class recovered in HPLC fractions 7-8 revealed little biological activity. HPLC fraction 20 dihydroceramides, substituted with 1-O-phosphoglycerol and isoC15:0 linked to the hydroxyl of 3-OH isoC17:0, significantly potentiated interleukin-1β (IL-1β) (-mediated prostaglandin secretion and produced marked alterations in fibroblast morphology. HPLC fraction 28 dihydroceramides, substituted with 1-O-phosphoethanolamine, demonstrated little capacity to potentiate IL-1β-mediated prostaglandin secretion. The novel phosphorylated dihydroceramides synthesized by P. gingivalis demonstrate varying biological activities based on the phosphorylated head group substitution and/or the addition of esterified fatty acid. These results also demonstrate the strong virulence capacity of phosphoglycerol dihydroceramides of P. gingivalis to promote inflammatory factor secretion from IL-1 β-treated fibroblasts and to produce marked alterations in cell morphology in culture.

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KW - Interleukin-1β

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KW - Phosphoethanolamine ceramide

KW - Phosphoglycerol dihydroceramide

KW - Prostaglandin E

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