出版社：American Society for Biochemistry and Molecular Biology
摘要：Porphyromonas 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 iso C17:0 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 iso C15:0 linked to the hydroxyl of 3-OH iso C17:0, 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-1β-treated fibroblasts and to produce marked alterations in cell morphology in culture.
