Further evidence that major outer membrane proteins homologous to OmpA in Porphyromonas gingivalis stabilize bacterial cells.
BACKGROUND
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.


CONCLUSIONS
This report provides further evidence that Pgm6/7 proteins in P. gingivalis play an important role in the maintenance of bacterial outer membrane integrity.
Iwami
J
J
Department of Microbiology, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi, Japan.
Murakami
Y
Y
Nagano
K
K
Nakamura
H
H
Yoshimura
F
F
D018829:Adhesins, Bacterial / Q000032:analysis; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000502:physiology*; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D003546:Cysteine Endopeptidases / Q000032:analysis; D022162:Cytoplasmic Vesicles / Q000648:ultrastructure; D017353:Gene Deletion; D000080867:Gingipain Cysteine Endopeptidases; D006801:Humans; D046529:Microscopy, Electron, Transmission; D009154:Mutation / Q000235:genetics; D016966:Porphyromonas gingivalis / Q000502:physiology* / Q000648:ultrastructure