| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
(Received for publication, September 27, 1994; and in revised form, November 18,
1994) From the
The identification of proteinases of Porphyromonas
gingivalis that act as virulence factors in periodontal disease
has important implications in the study of host-pathogen interactions
as well as in the discovery of potential therapeutic and
immunoprophylactic agents. We have cloned and characterized a gene that
encodes the 50-kDa cysteine proteinase gingipain or Arg-gingipain-1
(RGP-1) described previously (Chen, Z., Potempa, J., Polanowski, A.,
Wikstrom, M., and Travis, J. (1992) J. Biol. Chem. 267,
18896-18901). Analysis of the amino acid sequence of RGP-1
deduced from the cloned DNA sequence showed that the biosynthesis of
this proteinase involves processing of a polyprotein that contains
multiple adhesin molecules located at its carboxyl terminus. This
finding corroborates previous evidence (Pike R., McGraw, W., Potempa,
J., and Travis, J.(1994) J. Biol. Chem. 269, 406-411)
that RGP-1 is closely associated with adhesin molecules, and that high
molecular weight forms of the proteinase are involved in the binding of
erythrocytes. Mammalian periodontal diseases result from complex interactions
between the host and a variety of anaerobic microorganisms. A number of
studies have suggested an important role for Porphyromonas
gingivalis in human periodontal tissue
destruction(1, 2, 3) . Several potential
virulence factors, including the elaboration of proteinase activity,
have been identified for this
organism(4, 5, 6) . Proteinases have been
proposed to play a major role in periodontal disease because of their
capacity to degrade protective host immunoglobins and to hydrolyze host
proteins that provide amino acids required for growth, and by their
participation in the destruction of host connective tissue (7, 8, 9, 10, 11, 12, 13) .
Furthermore, there are reports indicating a direct involvement of
``trypsin-like'' proteinases of P. gingivalis in its
binding to erythrocytes and extracellular matrix components. This
suggests that some of the P. gingivalis proteolytic enzymes
associated with the cell surface function as adhesins that mediate
bacterial adherence to host tissues (reviewed in (14) ).
Several groups have reported the cloning of proteinase genes from P.
gingivalis(15, 16, 17, 18, 19, 20) .
We report here the molecular cloning of a gene that encodes gingipain-1
(RGP-1), ( Although the role of the RGP-1 proteinase in
the development of periodontal disease is not yet fully clear, recent
results have indicated that this proteinase is the major vascular
permeability enhancement factor of P. gingivalis, resulting in
gingival crevicular fluid production at sites of periodontitis caused
by infection with this organism(22) . It has been shown
previously that 110- and 95-kDa RGP-1 protein complexes possess
erythrocyte-binding properties(23) , suggesting an association
between proteolytic and hemolytic activities. Here we show, from
analysis of the amino acid sequence deduced from the rgp1 gene
sequence, that this proteinase/adhesin association is derived from the
biosynthesis of RGP-1 as a polyprotein that contains multiple adhesin
domains at the carboxyl terminus of the previously identified
proteinase. By comparison of the proteinase domain of the polyprotein
with the sequences of other cysteine proteinases, RGP-1 appears to be a
member of a new family of pathogenic proteinases.
We describe here that RGP-1, the major arginine-specific
cysteine proteinase from P. gingivalis, is synthesized as a
polyprotein that can function as an erythrocyte-binding protein through
the presence of multiple adhesin domains at its carboxyl terminus. Chen et al.(21) have determined the primary structure of
the amino terminus of RGP-1 by direct amino acid sequencing. This
sequence information was used to prepare a mixture of synthetic
oligonucleotides: primer GIN-1-32, a 32-mer coding for amino acids
2-8 of the mature protein (TPVEEKE); and primer GIN-2-30, a
30-mer coding for amino acids 25-32 of the mature protein
(KDFVDWKN). These primers were used to amplify from genomic DNA the
corresponding fragment of the rgp1 gene by PCR. The expected
105-base pair PCR product was cloned and sequenced. On the basis of
this sequence, GIN-8S-48, a unique 48-mer oligonucleotide probe
corresponding to the coding strand of rgp1, was synthesized
and used to screen a
Figure 1:
A, map of the cloned
genomic DNA sequence encompassing the rgp1 gene. Only major
restriction sites are indicated: B, BamHI; P, PstI; S, SmaI; A, Asp718; Pv, PvuII; H, HindIII. M13 subclones used for DNA sequencing are shown (arrows). The overlap at the 3`-PstI site was
determined by sequencing a SmaI/BamHI plasmid clone.
Also shown is a schematic representation of the RGP-1 polyprotein
structure, including the proposed methionine used for translation
initiation and experimentally determined basic residue cleavage sites.
Also shown (
The most striking feature of the deduced protein sequence is the
presence of multiple homologous sequences immediately carboxyl-terminal
to the proteinase coding domain (Fig. 1), leading to a
calculated molecular mass of 185.4 kDa of the encoded polyprotein.
Within these sequences can be found peptides identified by Pike et
al.(23) as the components of high molecular mass
gingipain (HGP) that confer adhesion activity on the high molecular
mass RGP-1 complex. The polyprotein sequence deduced from the gene
sequence now allows exact delineation of the primary structure of the
mature RGP-1 proteinase. The amino terminus (23) is derived
from proteolytic processing at an arginine residue (Fig. 1A). The carboxyl terminus is derived by
processing at Arg-492 and also releases the amino terminus of the
44-kDa HGP (HGP44, Fig. 1, A and B, underlined) determined by Pike et al.(23) .
Similar processing at Arg-1202 gives rise to the amino terminus of the
27-kDa HGP27 (Fig. 1, A and B, underlined) also found to be associated with RGP adhesion
activity (23) . More recently, we have used high resolution
SDS-PAGE (27) to separate the 95-kDa form of RGP into 5 major
bands of 50, 44, 27, 17, and 15 kDa. ( Three large repeats of homologous
sequence are located within three of the cleavage products (Fig. 2A). The first is found in the middle of HGP44,
the second in HGP17, and the third in the carboxyl-terminal region of
HGP27. Amino acid sequence identities within these 49-amino acid
stretches varies between 76 and 96%, indicating similar roles for each
sequence, possibly in non-covalent interactions with the proteinase
domain, or in the adhesion activity of the high molecular mass
complexes.
Figure 2:
A,
alignment of conserved areas within the adhesin domains of the RGP-1
polyprotein sequence. B, identification of the active site
cysteine residue as Cys-185 by active site labeling and separation of
V8 proteinase- and Asp-N-derived peptides. X refers to
unidentifiable amino acid residues at a given Edman degradation cycle.
During such analyses, these positions are characteristically associated
with cysteine or modified amino acid residues in the polypeptide
chain.
The mature RGP-1 sequence exhibited no similarity with
any cysteine proteinase reported previously except for the related
enzyme from the same organism, referred to as Lys-gingipain (23) . ( Molecular cloning of the rgp1 gene confirms previous
findings that this arginine-specific proteinase is closely associated
with adhesion activity. High trypsin-like activity has been shown
previously to be an important virulence factor in P.
gingivalis. Sequencing of the corresponding rgp1 gene of
the virulent W50 strain of P. gingivalis revealed only two
conservative amino acid changes in the mature enzyme sequence (data not
shown). Thus, any involvement of RGP-1 in virulence would have to be
due to its differential regulation, and enhanced expression in virulent
strains. The availability of RGP-1 DNA sequences will now allow the
further study of hemolysis of erythrocytes through the
adhesin/hemagglutinin activities of this proteolytic polyprotein.
Recombinant polypeptides can also be used for the development of
potential immunoprophylactic and therapeutic agents against this human
pathogen. The nucleotide
sequence(s) reported in this paper has been submitted to the
GenBank(TM)/EMBL Data Bank with accession number(s)
U15282[GenBank].
Volume 270,
Number 3,
Issue of January 20, 1995 pp. 1007-1010
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
BIOSYNTHESIS AS A PROTEINASE-ADHESIN POLYPROTEIN (*)
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS AND DISCUSSION
FOOTNOTES
ACKNOWLEDGEMENTS
REFERENCES
)a 50-kDa arginine-specific cysteine proteinase,
described previously by Chen et al.(21) , and found
predominantly in culture medium as 95- and 50-kDa proteins, or
associated with bacterial membranous fractions as 110- and
70-90-kDa forms.
Bacterial Strains
P. gingivalis strains
H66 and W50 were obtained from Dr. Roland Arnold, Emory University,
Atlanta, GA.Nucleic Acids
Oligonucleotides were synthesized by
the phosphoramidite method using an Applied Biosystems model 394 DNA
synthesizer, purified by PAGE, and desalted on Sep-Pak cartridges
(Millipore). DNA templates used for PCR were isolated from P.
gingivalis strain H66 by standard procedures(24) .Genomic Library Construction
A DASH P.
gingivalis H66 DNA library was constructed using the
DASH(TM) II/BamHI cloning kit (Stratagene). A library
of 2
10
independent recombinant clones was
obtained. ZAP P. gingivalis H66 and W50 DNA libraries
were also constructed as described using the
ZAP/BamHI
cloning kit (Stratagene). Libraries of 3
10 and 1.5
10 independent recombinant clones were obtained
from strains H66 and W50, respectively.Southern Blot Analysis
BamHI, HindIII, or PstI digestions of P. gingivalis H66 DNA were blotted (24) and hybridized with P-labeled oligonucleotide probes. Membranes were washed as
described below.
Screening of Genomic Libraries
Approximately
10
to 10 phage were grown on 5 150-mm
plates, lifted in duplicate onto supported nitrocellulose transfer
membranes (BAS-NC from Schleicher & Schuell). Hybridizations were
performed overnight at 42 °C in 2 Denhardt's
solution(24) , 6 SSC (SSC is 15 mM sodium
citrate, 150 mM NaCl), 0.4% SDS (w/v), 500 mg/ml salmon sperm
DNA. Filters were washed in 2 SSC containing 0.05% SDS (w/v) at
48 °C. Positively hybridizing plaques were purified. Standard
protocols for cDNA library screening, phage purification, agarose
gel electrophoresis, and plasmid cloning were used(24) . For
cloning of the 3`-region of the rgp1 gene, PstI/HindIII-digested DNA (50 µg) was
size-selected on 1% agarose, and the region at
4.5 kbp was cloned
into pBluescript SK(-). Positive clones were identified using a
20-mer oligonucleotide probe.
DNA Sequencing
Double-stranded DNA cloned into
pBluescript SK(-) and single-stranded DNA cloned into M13mp18 and
M13mp19 were sequenced by the dideoxy method(25) , using
sequencing kits purchased from United States Biochemicals (Sequenase
version 2.0). A 6327-base pair PstI/PvuII fragment
encoding the full RGP-1 polyprotein gene was submitted to GenBank under
the accession number U15282. Gene fragments obtained by PstI, PstI/Asp718, and BamHI/HindIII
digestion of plasmid subclones were introduced into M13 vectors, and
the sequence was obtained by single strand sequencing of M13 subclones
in both directions.Active Site Titration
RGP-1 from P. gingivalis H66 was purified and titrated as described
previously(21, 22, 23) .
Tosyl-L-lysine chloromethyl ketone (TLCK) and sequencing grade
enzymes: Staphylococcus aureus V8 protease (Glu-C) and Asp-N
endopeptidase were from Boehringer Mannheim. Bio-X-FPRck was purchased
from Haematologic Technologies Inc. (Essex Jct., VT), and avidin
(monomeric)-agarose was obtained from Sigma.Active Site Labeling
RGP-1 (6.4 µM)
was activated in 75 mM HEPES, 2 mM CaCl
,
8 mM cysteine, pH 8.0, for 15 min at 37 °C, then treated
with a 1.2 molar excess of Bio-X-FPRck and incubated for 15 min at room
temperature. Residual RGP activity (less than 5%) was eliminated by
treatment with TLCK (final concentration 2 mM), the sample was
dialyzed extensively against 50 mM ammonium bicarbonate, pH
7.8 and lyophilized. Biotinylated RGP (50 nmol) was denatured in 6 M guanidine HCl, reduced with dithiothreitol, and S-carboxymethylated by the method of Waxdal et
al.(26) . The sample was desalted on a PD-10 column
(Pharmacia Biotech Inc.) equilibrated with 50 mM ammonium
bicarbonate pH 7.8, and the protein concentration determined by the BCA
method (Pierce).Polypeptide Chain Fragmentation and Analysis
The
derivatized protein (25 nmol) was digested in 25 mM ammonium
bicarbonate buffer pH 7.8 with V8 protease at 25 °C for 12 h (1:100
enzyme to substrate weight ratio) or with Asp-N endopeptidase at 37
°C for 10 h (1:1000 enzyme to substrate weight ratio). Each digest
(2 ml) was loaded on a 5-ml avidin-agarose column equilibrated with 0.1 M Tris, pH 8.0. The column was washed with 30 ml of the
equilibration buffer, followed by 30 ml of 1.0 M NaCl/Tris, pH
8.0, then 50 ml of deionized HO. Biotinylated peptides were
eluted with HCl (pH 2.0), and 5-ml fractions were collected directly
into 1 ml of 0.2 M ammonium bicarbonate. Biotinylated
peptide-containing fractions were detected by dot blot analysis on
nitrocellulose membranes. Blotted membranes were incubated with
streptavidin-alkaline phosphatase conjugate and developed with the
nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate reagent kit
from Bio-Rad. The fractions that contained biotinylated peptide were
pooled, blotted onto polyvinylidine difluoride membranes, and subjected
to amino-terminal amino acid sequence analysis using an Applied
Biosystems 460A gas phase sequencer.
DASH DNA library constructed from BamHI-digested P. gingivalis genomic DNA. DNA
sequence analysis of positive clones indicated that the proteinase
domain was encoded by these
3.5-kbp clones. However, since no
transcriptional termination codon was evident within the large open
reading frame encoding the proteinase, overlapping clones were isolated
from a size-selected PstI/HindIII plasmid library,
using a 20-mer oligonucleotide probe. Using this procedure, several
4.5-kbp-containing clones were obtained. In total,
7.8 kbp of
genomic DNA from BamHI to HindIII sites (Fig. 1A) was isolated and characterized. The composite
6327-base pair PstI/PvuII fragment of this genomic
DNA (Fig. 1A) was fully sequenced in both directions
and is described here, with the first base of the 5`-PstI site (Fig. 1A) assigned as base 1. Within this composite
sequence was found an open reading frame encoding a 1704 amino acid
sequence (Fig. 1B), with the 5`-most ATG initiation
codon at nucleotides 949-951. Between this ATG and the mature
RGP-1 sequence are an additional 8 in-frame methionine codons. The
exact ATG used for initiation of translation is currently unknown,
although the presence of a consensus TATA box (TATAAT) at nucleotides
889-894 suggests the 5`-most ATG as the strongest candidate.
) is the experimentally determined active site
cysteine residue of the previously identified
50-kDa mature
RGP-1(21) . B, full deduced amino acid sequence of the
RGP-1 polyprotein in single-letter code. Peptide sequences
identified previously(23) , or as part of the present work, are underlined. The first amino acid of the mature RGP-1
proteinase (tyrosine) is assigned as amino acid
1.
)Amino-terminal
sequence analysis confirmed the structures of the 50-, 44-, and 27-kDa
fragments reported previously(23) . For the 17- and 15-kDa
fragments, the following amino termini were determined: PQSVWIERTVDL
and ADFTETFESSTHG, respectively. Thus, the 17-kDa polypeptide (HGP17, Fig. 1, A and B, underlined) is
cleaved at lysine residue 1044, most likely catalyzed by Lys-gingipain,
the other cysteine proteinase produced in large quantities by P.
gingivalis(23) . The sequence of the 15-kDa fragment
(HGP15, Fig. 1, A and B, underlined)
reveals that processing occurs after Arg-909. The calculated molecular
mass of 53.9 kDa for RGP-1 is in good agreement with its mobility of
50 kDa on SDS-PAGE. Similarly, the 417-, 275-, 158-, and 135-amino
acid sequences of HGP44 (44.7 kDa), HGP27 (29.6 kDa), HGP17 (17.5 kDa),
and HGP15 (14.3 kDa), respectively, correlate well with their SDS-PAGE
mobilities. Together, the proteinase domain and adhesin/hemagglutinin
fragments would create a polyprotein of 159.9 kDa, while the high
molecular mass form of RGP (HGP) is only 95 kDa, indicating that the
secreted enzyme is most likely processed and assembled as a
non-covalent complex of the proteinase with different individual
adhesin/hemagglutinin domains.
)Even in this case, the similarity is
limited to the sequence around His-211 and Asn-442, suggesting that
these residues, along with the active site cysteine residue (Cys-185),
determined by active site labeling (Fig. 2B), encompass
the catalytic triad. Interestingly, the lack of any similarity around
these sequences with cysteine proteinases other than Lys-gingipain
suggests that these bacterial proteinases represent a distinct branch
of this family of proteolytic enzymes. That the catalytic apparatus of
RGP-1 might be different from that of other known cysteinyl proteinases
demands that residues other than Cys-185 involved directly in the
hydrolysis of peptide bonds must be verified experimentally.
)-aminocaproyl-Phe-Pro-Arg-chloromethylketone; kbp, kilobase
pair(s); HGP, high molecular mass gingipain; PCR, polymerase chain
reaction; TLCK, tosyl-L-lysine chloromethyl ketone.
))
We gratefully acknowledge the technical contributions
of W.-C. A. Chen, T. Rigley, and K. Norris.
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  E. Sakai, M. Naito, K. Sato, H. Hotokezaka, T. Kadowaki, A. Kamaguchi, K. Yamamoto, K. Okamoto, and K. Nakayama Construction of Recombinant Hemagglutinin Derived from the Gingipain-Encoding Gene of Porphyromonas gingivalis, Identification of Its Target Protein on Erythrocytes, and Inhibition of Hemagglutination by an Interdomain Regional Peptide J. Bacteriol., June 1, 2007; 189(11): 3977 - 3986. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Yamatake, M. Maeda, T. Kadowaki, R. Takii, T. Tsukuba, T. Ueno, E. Kominami, S. Yokota, and K. Yamamoto Role for Gingipains in Porphyromonas gingivalis Traffic to Phagolysosomes and Survival in Human Aortic Endothelial Cells Infect. Immun., May 1, 2007; 75(5): 2090 - 2100. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. D. Pathirana, N. M. O'Brien-Simpson, G. C. Brammar, N. Slakeski, and E. C. Reynolds Kgp and RgpB, but Not RgpA, Are Important for Porphyromonas gingivalis Virulence in the Murine Periodontitis Model Infect. Immun., March 1, 2007; 75(3): 1436 - 1442. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. A. Seers, N. Slakeski, P. D. Veith, T. Nikolof, Y.-Y. Chen, S. G. Dashper, and E. C. Reynolds The RgpB C-Terminal Domain Has a Role in Attachment of RgpB to the Outer Membrane and Belongs to a Novel C-Terminal-Domain Family Found in Porphyromonas gingivalis. J. Bacteriol., September 1, 2006; 188(17): 6376 - 6386. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Sato, E. Sakai, P. D. Veith, M. Shoji, Y. Kikuchi, H. Yukitake, N. Ohara, M. Naito, K. Okamoto, E. C. Reynolds, et al. Identification of a New Membrane-associated Protein That Influences Transport/Maturation of Gingipains and Adhesins of Porphyromonas gingivalis J. Biol. Chem., March 11, 2005; 280(10): 8668 - 8677. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Takii, T. Kadowaki, A. Baba, T. Tsukuba, and K. Yamamoto A Functional Virulence Complex Composed of Gingipains, Adhesins, and Lipopolysaccharide Shows High Affinity to Host Cells and Matrix Proteins and Escapes Recognition by Host Immune Systems Infect. Immun., February 1, 2005; 73(2): 883 - 893. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K.-A. Nguyen, A. A. DeCarlo, M. Paramaesvaran, C. A. Collyer, D. B. Langley, and N. Hunter Humoral Responses to Porphyromonas gingivalis Gingipain Adhesin Domains in Subjects with Chronic Periodontitis Infect. Immun., March 1, 2004; 72(3): 1374 - 1382. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M.-P. Savariau-Lacomme, C. Lebarbier, T. Karjalainen, A. Collignon, and C. Janoir Transcription and Analysis of Polymorphism in a Cluster of Genes Encoding Surface-Associated Proteins of Clostridium difficile J. Bacteriol., August 1, 2003; 185(15): 4461 - 4470. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Kamaguchi, T. Ohyama, E. Sakai, R. Nakamura, T. Watanabe, H. Baba, and K. Nakayama Adhesins encoded by the gingipain genes of Porphyromonas gingivalis are responsible for co-aggregation with Prevotella intermedia Microbiology, May 1, 2003; 149(5): 1257 - 1264. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Mikolajczyk, K. M. Boatright, H. R. Stennicke, T. Nazif, J. Potempa, M. Bogyo, and G. S. Salvesen Sequential Autolytic Processing Activates the Zymogen of Arg-gingipain J. Biol. Chem., March 14, 2003; 278(12): 10458 - 10464. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. L. W. Yun, A. A. DeCarlo, C. Collyer, and N. Hunter Modulation of an Interleukin-12 and Gamma Interferon Synergistic Feedback Regulatory Cycle of T-Cell and Monocyte Cocultures by Porphyromonas gingivalis Lipopolysaccharide in the Absence or Presence of Cysteine Proteinases Infect. Immun., October 1, 2002; 70(10): 5695 - 5705. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Shoji, D. B. Ratnayake, Y. Shi, T. Kadowaki, K. Yamamoto, F. Yoshimura, A. Akamine, M. A. Curtis, and K. Nakayama Construction and characterization of a nonpigmented mutant of Porphyromonas gingivalis: cell surface polysaccharide as an anchorage for gingipains Microbiology, April 1, 2002; 148(4): 1183 - 1191. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. A. Burgess, D. F. Kirke, P. Williams, K. Winzer, K. R. Hardie, N. L. Meyers, J. Aduse-Opoku, M. A. Curtis, and M. Camara LuxS-dependent quorum sensing in Porphyromonas gingivalis modulates protease and haemagglutinin activities but is not essential for virulence Microbiology, March 1, 2002; 148(3): 763 - 772. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. P. C. A. Lima, P. C. Almeida, I. L. S. Tersariol, V. Schmitz, A. H. Schmaier, L. Juliano, I. Y. Hirata, W. Muller-Esterl, J. R. Chagas, and J. Scharfstein Heparan Sulfate Modulates Kinin Release by Trypanosoma cruzi through the Activity of Cruzipain J. Biol. Chem., February 15, 2002; 277(8): 5875 - 5881. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. C. Fenno, S. Y. Lee, C. H. Bayer, and Y. Ning The opdB Locus Encodes the Trypsin-Like Peptidase Activity of Treponema denticola Infect. Immun., October 1, 2001; 69(10): 6193 - 6200. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Olczak, D. W. Dixon, and C. A. Genco Binding Specificity of the Porphyromonas gingivalis Heme and Hemoglobin Receptor HmuR, Gingipain K, and Gingipain R1 for Heme, Porphyrins, and Metalloporphyrins J. Bacteriol., October 1, 2001; 183(19): 5599 - 5608. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Sroka, M. Sztukowska, J. Potempa, J. Travis, and C. A. Genco Degradation of Host Heme Proteins by Lysine- and Arginine-Specific Cysteine Proteinases (Gingipains) of Porphyromonas gingivalis J. Bacteriol., October 1, 2001; 183(19): 5609 - 5616. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. L. W. Yun, A. A. Decarlo, C. Collyer, and N. Hunter Hydrolysis of Interleukin-12 by Porphyromonas gingivalis Major Cysteine Proteinases May Affect Local Gamma Interferon Accumulation and the Th1 or Th2 T-Cell Phenotype in Periodontitis Infect. Immun., September 1, 2001; 69(9): 5650 - 5660. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Oido-Mori, R. Rezzonico, P.-L. Wang, Y. Kowashi, J.-M. Dayer, P. C. Baehni, and C. Chizzolini Porphyromonas gingivalis Gingipain-R Enhances Interleukin-8 but Decreases Gamma Interferon-Inducible Protein 10 Production by Human Gingival Fibroblasts in Response to T-Cell Contact Infect. Immun., July 1, 2001; 69(7): 4493 - 4501. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Yonezawa, K. Ishihara, and K. Okuda Arg-Gingipain A DNA Vaccine Induces Protective Immunity against Infection by Porphyromonas gingivalis in a Murine Model Infect. Immun., May 1, 2001; 69(5): 2858 - 2864. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Chen, K. Nakayama, L. Belliveau, and M. J. Duncan Porphyromonas gingivalis Gingipains and Adhesion to Epithelial Cells Infect. Immun., May 1, 2001; 69(5): 3048 - 3056. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Rice, R. Peralta, D. Bast, J. de Azavedo, and M. J. McGavin Description of Staphylococcus Serine Protease (ssp) Operon in Staphylococcus aureus and Nonpolar Inactivation of sspA-Encoded Serine Protease Infect. Immun., January 1, 2001; 69(1): 159 - 169. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Aduse-Opoku, N. N. Davies, A. Gallagher, A. Hashim, H. E. A. Evans, M. Rangarajan, J. M. Slaney, and M. A. Curtis Generation of Lys-gingipain protease activity in Porphyromonas gingivalis W50 is independent of Arg-gingipain protease activities Microbiology, August 1, 2000; 146(8): 1933 - 1940. [Abstract] [Full Text]
|
|
|
|
|
|
N. M. O'Brien-Simpson, R. A. Paolini, and E. C. Reynolds RgpA-Kgp Peptide-Based Immunogens Provide Protection against Porphyromonas gingivalis Challenge in a Murine Lesion Model Infect. Immun., July 1, 2000; 68(7): 4055 - 4063. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. M. O'Brien-Simpson, C. L. Black, P. S. Bhogal, S. M. Cleal, N. Slakeski, T. J. Higgins, and E. C. Reynolds Serum Immunoglobulin G (IgG) and IgG Subclass Responses to the RgpA-Kgp Proteinase-Adhesin Complex of Porphyromonas gingivalis in Adult Periodontitis Infect. Immun., May 1, 2000; 68(5): 2704 - 2712. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Kozarov, N. Miyashita, J. Burks, K. Cerveny, T. A. Brown, W. P. McArthur, and A. Progulske-Fox Expression and Immunogenicity of Hemagglutinin A from Porphyromonas gingivalis in an Avirulent Salmonella enterica Serovar Typhimurium Vaccine Strain Infect. Immun., February 1, 2000; 68(2): 732 - 739. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Dong, T. Chen, F. E. Dewhirst, R. D. Fleischmann, C. M. Fraser, and M. J. Duncan Genomic Loci of the Porphyromonas gingivalis Insertion Element IS1126 Infect. Immun., July 1, 1999; 67(7): 3416 - 3423. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Shi, D. B. Ratnayake, K. Okamoto, N. Abe, K. Yamamoto, and K. Nakayama Genetic Analyses of Proteolysis, Hemoglobin Binding, and Hemagglutination of Porphyromonas gingivalis. CONSTRUCTION OF MUTANTS WITH A COMBINATION OF rgpA, rgpB, kgp, AND hagA J. Biol. Chem., June 18, 1999; 274(25): 17955 - 17960. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. L. W. Yun, A. A. DeCarlo, and N. Hunter Modulation of Major Histocompatibility Complex Protein Expression by Human Gamma Interferon Mediated by Cysteine Proteinase-Adhesin Polyproteins of Porphyromonas gingivalis Infect. Immun., June 1, 1999; 67(6): 2986 - 2995. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. P. Lewis and F. L. Macrina Localization of HArep-Containing Genes on the Chromosome of Porphyromonas gingivalis W83 Infect. Immun., May 1, 1999; 67(5): 2619 - 2623. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Shibata, M. Hayakawa, H. Takiguchi, T. Shiroza, and Y. Abiko Determination and Characterization of the Hemagglutinin-associated Short Motifs Found in Porphyromonas gingivalis Multiple Gene Products J. Biol. Chem., February 19, 1999; 274(8): 5012 - 5020. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. A. Barkocy-Gallagher, J. W. Foley, and M. S. Lantz Activities of the Porphyromonas gingivalis PrtP Proteinase Determined by Construction of prtP-Deficient Mutants and Expression of the Gene in Bacteroides Species J. Bacteriol., January 1, 1999; 181(1): 246 - 255. [Abstract] [Full Text]
|
|
|
|
 |
|
 R. J. Lamont and H. F. Jenkinson Life Below the Gum Line: Pathogenic Mechanisms of Porphyromonas gingivalis Microbiol. Mol. Biol. Rev., December 1, 1998; 62(4): 1244 - 1263. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Kadowaki, K. Nakayama, F. Yoshimura, K. Okamoto, N. Abe, and K. Yamamoto Arg-gingipain Acts as a Major Processing Enzyme for Various Cell Surface Proteins in Porphyromonas gingivalis J. Biol. Chem., October 30, 1998; 273(44): 29072 - 29076. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
|
