INTRODUCTION: Porphyromonas gingivalis is one of the most important bacteria in the progression of chronic periodontal disease. We hypothesized that the major outer membrane proteins Pgm6/7, which are homologous to the OmpA protein in Escherichia coli, might contribute to the stabilization of the cell surface. In this study, the effects of Pgm6/7 on the cell surface were examined morphologically. METHODS: Deletion mutants of Pgm6/7 (Delta694, Delta695 and Delta695-694) were constructed using the polymerase chain reaction-based overlap extension method. Wild-type ATCC 33277 and Pgm6/7 mutants were grown under anaerobic conditions. Whole cells and thin sections of fixed cells were stained and examined by transmission electron microscopy. RESULTS: Compared with the wild-type, numerous vesicles released from cells were observed in each deletion mutant. The outer membrane appeared wavy and irregular. Increased numbers of vesicles were confirmed after their preparation from the culture supernatant. Total gingipain activity in vesicles was increased five- to 10-fold in the deletion mutants. CONCLUSION: This report provides further evidence that Pgm6/7 proteins in P. gingivalis play an important role in the maintenance of bacterial outer membrane integrity.
INTRODUCTION:Porphyromonas gingivalis is one of the most important bacteria in the progression of chronic periodontal disease. We hypothesized that the major outer membrane proteins Pgm6/7, which are homologous to the OmpA protein in Escherichia coli, might contribute to the stabilization of the cell surface. In this study, the effects of Pgm6/7 on the cell surface were examined morphologically. METHODS: Deletion mutants of Pgm6/7 (Delta694, Delta695 and Delta695-694) were constructed using the polymerase chain reaction-based overlap extension method. Wild-type ATCC 33277 and Pgm6/7 mutants were grown under anaerobic conditions. Whole cells and thin sections of fixed cells were stained and examined by transmission electron microscopy. RESULTS: Compared with the wild-type, numerous vesicles released from cells were observed in each deletion mutant. The outer membrane appeared wavy and irregular. Increased numbers of vesicles were confirmed after their preparation from the culture supernatant. Total gingipain activity in vesicles was increased five- to 10-fold in the deletion mutants. CONCLUSION: This report provides further evidence that Pgm6/7 proteins in P. gingivalis play an important role in the maintenance of bacterial outer membrane integrity.
Authors: Jacqueline E Heath; Christine A Seers; Paul D Veith; Catherine A Butler; Nor A Nor Muhammad; Yu-Yen Chen; Nada Slakeski; Benjamin Peng; Lianyi Zhang; Stuart G Dashper; Keith J Cross; Steven M Cleal; Caroline Moore; Eric C Reynolds Journal: PLoS One Date: 2016-10-06 Impact factor: 3.240
Authors: Kathryn L Naylor; Magdalena Widziolek; Stuart Hunt; Mary Conolly; Matthew Hicks; Prachi Stafford; Jan Potempa; Craig Murdoch; C W Ian Douglas; Graham P Stafford Journal: Microbiologyopen Date: 2016-09-06 Impact factor: 3.139