Roles of porphyrins and host iron transport proteins in regulation of growth of Porphyromonas gingivalis W50.
Porphyromonas gingivalis (Bacteroides gingivalis) requires iron in the form of hemin for growth and virulence in vitro, but the contributions of the porphyrin ring structure, porphyrin-associated iron, host hemin-sequestering molecules, and host iron-withholding proteins to its survival are unknown. Therefore, the effects of various porphyrins, host iron transport proteins, and inorganic iron sources on the growth of P. gingivalis W50 were examined to delineate the various types of iron molecules used for cellular metabolism. Cell envelope-associated hemin and iron stores contributed to the growth of P. gingivalis in hemin-free culture, and depletion of these endogenous reserves required eight serial transfers into hemin-free medium for total suppression of growth. Comparable growth of P. gingivalis was observed with 7.7 microM equivalents of hemin as hemoglobin (HGB), methemoglobin, myoglobin, hemin-saturated serum albumin, lactoperoxidase, cytochrome c, and catalase. Unrestricted growth was recorded in the presence of haptoglobin-HGB and hemopexin-hemin complexes, indicating that these host defense proteins do not sequester HGB and hemin from P. gingivalis. The iron chelator 2,2'-bipyridyl functionally chelated hemin-associated iron, resulting in dose-dependent inhibition of growth in hemin-restricted cultures at 1 to 25 microM 2,2'-bipyridyl concentrations. In the absence of an exogenous iron source, protoporphyrin IX did not support P. gingivalis growth. These findings suggest that the iron atom in the hemin molecule is the critical constituent for growth and that the tetrapyrrole porphyrin ring structure may represent an important vehicle for delivery of iron into the P. gingivalis cell. P. gingivalis does not have a strict requirement for porphyrins, since growth occurred with nonhemin iron sources, including high concentrations (200 muM) of ferric, ferrous, and nitrogenous inorganic iron, and P. gingivalis exhibited unrestricted growth in the presence of host transferrin, lactoferrin, and serum albumin. The diversity of iron substrates utilized by P. gingivalis and the observation that growth was not affected by the bacteriostatic effects of host iron-withholding proteins, which it may encounter in the periodontal pocket, may explain why P. gingivalis is such a formidable pathogen in the periodontal disease process.
Bramanti
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University of Texas Health Science Center, San Antonio 78284-7894.
Holt
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D000818:Animals; D001798:Blood Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D002374:Catalase / Q000494:pharmacology; D002417:Cattle; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology; D003574:Cytochrome c Group / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development; D006418:Heme / Q000378:metabolism* / Q000494:pharmacology; D006454:Hemoglobins / Q000302:isolation & purification / Q000494:pharmacology*; D006736:Horses; D006801:Humans; D007501:Iron / Q000494:pharmacology*; D007784:Lactoperoxidase / Q000494:pharmacology; D008706:Methemoglobin / Q000494:pharmacology; D009211:Myoglobin / Q000302:isolation & purification / Q000494:pharmacology; D011166:Porphyrins / Q000494:pharmacology*; D016966:Porphyromonas gingivalis / Q000187:drug effects / Q000254:growth & development* / Q000472:pathogenicity; D012709:Serum Albumin / Q000302:isolation & purification / Q000494:pharmacology; D014774:Virulence / Q000187:drug effects / Q000502:physiology*
D001798:Blood Proteins; D003574:Cytochrome c Group; D006454:Hemoglobins; D009211:Myoglobin; D011166:Porphyrins; D012709:Serum Albumin; D006418:Heme; D008706:Methemoglobin; D007501:Iron; D007784:Lactoperoxidase; D002374:Catalase