Characterization and expression of a novel Porphyromonas gingivalis outer membrane protein, Omp28.
We report the characterization of a Porphyromonas gingivalis gene, designated omp28, encoding a protein that we have previously purified and characterized as a 28-kDa outer membrane protein. The deduced amino acid sequence of the omp28 open reading frame displayed an outer membrane leader sequence and lipoprotein attachment site but did not exhibit any significant overall sequence identity with protein sequences in the databases. A small stretch of amino acids (19 residues) exhibits 50% sequence identity with a segment of a fimbrial protein from Dichelobacter nodosus involved in adhesion, suggesting that Omp28 may be a surface adhesin/receptor of P. gingivalis. Using the pET-24 vector we expressed recombinant Omp28 (rOmp28) in Escherichia coli. Western blot analyses of purified rOmp28 with rabbit antisera to a P. gingivalis outer membrane preparation, protective rat anti-whole P. gingivalis antisera and pooled human sera from chronic periodontitis patients showed that the recombinant was recognized by all antisera. Further, anti-rOmp28 antisera exhibited strong reactivity with a panel of four laboratory strains and 10 clinical isolates of P. gingivalis from the United States, Sudan, Romania and Norway. These results suggest that Omp28 is expressed by a wide distribution of P. gingivalis strains.
Slakeski
N
N
School of Dental Science, The University of Melbourne, 711 Elizabeth Street, Victoria 3000, Australia.
Margetts
M
M
Moore
C
C
Czajkowski
L
L
Barr
I G
IG
Reynolds
E C
EC
D018829:Adhesins, Bacterial / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D001665:Binding Sites / Q000235:genetics; D015139:Blotting, Southern; D015153:Blotting, Western; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D020376:Dichelobacter nodosus / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008074:Lipoproteins / Q000235:genetics; D016366:Open Reading Frames / Q000235:genetics; D010518:Periodontitis / Q000097:blood; D016966:Porphyromonas gingivalis / Q000235:genetics* / Q000276:immunology; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020539:Sequence Analysis, Protein
D018829:Adhesins, Bacterial; D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D004274:DNA, Recombinant; D008074:Lipoproteins; C116185:OMP-28 protein, Salmonella typhi