Purification and characterization of fibroblast-activating factor isolated from Porphyromonas gingivalis W50.
A 24-kDa polypeptide which activated the incorporation of [3H]thymidine into human fibroblasts was isolated from the outer membrane vesicles of Porphyromonas gingivalis W50. This polypeptide, named fibroblast activating factor (FAF), was isolated by 3-[(3-cholamidopropyl)-dimethyl-ammonio]-1-propane-sulfonate (CHAPS) detergent extraction and purified by DEAE ion-exchange chromatography and preparative isoelectric focusing. Purified FAF (100 ng of protein per ml) caused a 400% increase in [3H]thymidine incorporation into human gingival fibroblasts (HGFs) compared with results for controls. FAF was characterized as (i) a polypeptide with molecular masses of 24 kDa when heated at 100 degrees C for 5 min and 44 kDa when unheated, (ii) heat sensitive but not affected by selected reducing reagents, and (iii) possessing slight phosphatase activity. N'-terminal sequence analysis revealed no homology with P. gingivalis peptides or with any host-derived growth factors. These data suggest that FAF functions as a significant virulence factor which in vivo is capable of modulating homeostasis in local connective tissues.
Department of Periodontics, University of Texas Health Science Center, San Antonio 78284-7894.
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Mihara
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SC
Holt
D000595:Amino Acid Sequence; D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D002455:Cell Division / Q000187:drug effects; D010450:Endopeptidases; D005347:Fibroblasts / Q000187:drug effects; D018093:Gelatinases; D006133:Growth Substances / Q000302:isolation & purification* / Q000494:pharmacology; D006801:Humans; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016966:Porphyromonas gingivalis / Q000737:chemistry*; D012697:Serine Endopeptidases; D000099167:Fibroblast Activation Protein Alpha
D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D010450:Endopeptidases; D018093:Gelatinases; D006133:Growth Substances; D008565:Membrane Proteins; D012697:Serine Endopeptidases; D000099167:Fibroblast Activation Protein Alpha