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Infect Immun, March 1998, p. 1159-1166, Vol. 66, No. 3
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Role of Arg-Gingipain A in Virulence of Porphyromonas gingivalis

Masayuki Tokuda,1,dagger Thonthi Karunakaran,1 Margaret Duncan,2 Nobushiro Hamada,1,Dagger and Howard Kuramitsu1,*

Department of Oral Biology, State University of New York, Buffalo, New York 14214,1 and Forsyth Dental Center, Boston, Massachusetts 021152

Received 8 August 1997/Returned for modification 10 November 1997/Accepted 5 December 1997

In order to access the role of the Porphyromonas gingivalis Arg-gingipain proteases in the virulence of this organism, a mutant defective in the rgpA gene was constructed in strain 381. This mutant, MT10, displayed only 40% of the Arg-specific cysteine protease activity of the wild-type strain. In addition, MT10, as well as the recently characterized protease mutant G-102, which is defective in the rgpB gene, displayed reduced self-aggregation, hemagglutination, and the ability to bind to immobilized type I collagen compared to levels of the wild-type parent. However, unlike mutant G-102, the rgpA mutant displayed increased binding to epithelial cells relative to that of the parental organism. Mutant MT10 also did not express detectable levels of the FimA protein as assessed by both Western and Northern blotting or fimbriae visible by electron microscopy of the cells. Furthermore, the ability of MT10 to degrade rat tail collagen fibers when it was cultured at 37°C was markedly attenuated compared to that of strain 381. These results suggest that Arg-gingipain A may play a significant role in the pathogenicity of P. gingivalis by altering the colonization and toxic properties of the organism.

* Corresponding author. Mailing address: Department of Oral Biology, State University of New York, Buffalo, NY 14214. Phone: (716) 829-2068. Fax: (716) 829-3942. E-mail: Kuramits{at}ACSU.BUFFALO.EDU.

dagger Present address: Department of Operative Dentistry and Endodontology, Kagoshima University Dental School, Kagoshima, Japan.

Dagger Present address: Department of Oral Microbiology, Kanagawa Dental College, Yokosuka, Japan.




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