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PMID: 23264054 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Identification and characterization of Porphyromonas gingivalis client proteins that bind to Streptococcus oralis glyceraldehyde-3-phosphate dehydrogenase.

Infection and immunity ·Vol. 81 ·No. 3 ·2013-03-00 ·Pages 753-63

Maeda K, Nagata H, Kuboniwa M, Ojima M, Osaki T, Minamino N, Amano A

Abstract

Coaggregation of Porphyromonas gingivalis and oral streptococci is thought to play an important role in P. gingivalis colonization. Previously, we reported that P. gingivalis major fimbriae interacted with Streptococcus oralis glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and that amino acid residues 166 to 183 of GAPDH exhibited strong binding activity toward P. gingivalis fimbriae (H. Nagata, M. Iwasaki, K. Maeda, M. Kuboniwa, E. Hashino, M. Toe, N. Minamino, H. Kuwahara, and S. Shizukuishi, Infect. Immun. 77:5130-5138, 2009). The present study aimed to identify and characterize P. gingivalis components other than fimbriae that interact with S. oralis GAPDH. A pulldown assay was performed to detect potential interactions between P. gingivalis client proteins and S. oralis recombinant GAPDH with amino acid residues 166 to 183 deleted by site-directed mutagenesis. Seven proteins, namely, tonB-dependent receptor protein (RagA4), arginine-specific proteinase B, 4-hydroxybutyryl-coenzyme A dehydratase (AbfD), lysine-specific proteinase, GAPDH, NAD-dependent glutamate dehydrogenase (GDH), and malate dehydrogenase (MDH), were identified by two-dimensional gel electrophoresis followed by proteomic analysis using tandem mass spectrometry. Interactions between these client proteins and S. oralis GAPDH were analyzed with a biomolecular interaction analysis system. S. oralis GAPDH showed high affinity for five of the seven client proteins (RagA4, AbfD, GAPDH, GDH, and MDH). Interactions between P. gingivalis and S. oralis were measured by a turbidimetric method and fluorescence microscopy. RagA4, AbfD, and GDH enhanced coaggregation, whereas GAPDH and MDH inhibited coaggregation. Furthermore, the expression of luxS in P. gingivalis was upregulated by RagA4, AbfD, and GDH but was downregulated by MDH. These results indicate that the five P. gingivalis client proteins function as regulators in P. gingivalis biofilm formation with oral streptococci.

MeSH Terms
Bacterial Proteins/genetics,metabolism Biofilms Electrophoresis, Gel, Two-Dimensional Gene Expression Regulation, Bacterial/physiology Gene Expression Regulation, Enzymologic/physiology Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)/genetics,metabolism Immunoblotting Multienzyme Complexes/genetics,metabolism Mutation NADH, NADPH Oxidoreductases/genetics,metabolism Porphyromonas gingivalis/genetics,metabolism Protein Binding Streptococcus oralis/enzymology
Chemicals
Bacterial Proteins Multienzyme Complexes Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) NADH oxidase NADH, NADPH Oxidoreductases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Maeda Kazuhiko
Department of Preventive Dentistry, Osaka University Graduate School of Dentistry, Suita, Osaka, Japan. kmaeda@dent.osaka-u.ac.jp
Nagata Hideki
Kuboniwa Masae
Ojima Miki
Osaki Tsukasa
Minamino Naoto
Amano Atsuo
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
1098-5522
DOI
10.1128/IAI.00875-12
Email
kmaeda@dent.osaka-u.ac.jp
Published
2013-03-00
Epub
2012-00-21
Pages
753-63
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC3584861
Subset
IM
External Links
PubMed source DOI: 10.1128/IAI.00875-12 PMC Full Text View journal details
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