Abstract
The gingival seal around teeth prevents bacteria from destroying the tooth-supporting tissues and disseminating throughout the body. Porphyromonas gingivalis, a major periodontopathogen, degrades components of the specialized extracellular matrix that mediates attachment of the gingiva to the tooth. Of these, secretory calcium-binding phosphoprotein proline-glutamine rich 1 (SCPPPQ1) protein has a distinctive resistance to degradation, suggesting that it may offer resistance to bacterial attack. In silico analysis of its amino acid sequence was used to explore its molecular characteristics and to predict its two- and three-dimensional structure. SCPPPQ1 exhibits similarities with both proline-rich and cationic antimicrobial proteins, suggesting a putative antimicrobial potential. A combination of imaging approaches showed that incubation with 20 μM of purified SCPPPQ1 decrease bacterial number (p < 0.01). Fluorescence intensity decreased by 70% following a 2 h incubation of Porphyromonas gingivalis with the protein. Electron microscopy analyses revealed that SCPPPQ1 induced bacterial membrane disruption and breaches. While SCPPPQ1 has no effect on mammalian cells, our results suggest that it is bactericidal to Porphyromonas gingivalis, and that this protein, normally present in the gingival seal, may be exploited to maintain a healthy seal and prevent systemic dissemination of bacteria.
Highlights
The gingival seal around teeth prevents bacteria from destroying the tooth-supporting tissues and disseminating throughout the body
The results showed that rat SCPPPQ1 has few similarities with other proteins with the exception of Bactenecin 7 (Bac-7), a well-known prolinerich AMPs (PrAMPs) produced by bovine n eutrophils[25]
The objective of our work was to better characterize the antibacterial potential of rat SCPPPQ1 protein, a structural matrix molecule produced by the junctional epithelium (JE) that is resistant to bacterial degradation[13]
Summary
The gingival seal around teeth prevents bacteria from destroying the tooth-supporting tissues and disseminating throughout the body. While SCPPPQ1 has no effect on mammalian cells, our results suggest that it is bactericidal to Porphyromonas gingivalis, and that this protein, normally present in the gingival seal, may be exploited to maintain a healthy seal and prevent systemic dissemination of bacteria. A specialized portion of the gingiva, called junctional epithelium (JE), seals off the tooth supporting tissues from the aggressive environment of the oral c avity[11] Under healthy conditions, it prevents the local infiltration of bacteria as well as their dissemination throughout the b ody[5]. The SCPP gene cluster encodes for a number of proteins possessing antimicrobial properties[21], including some well-known AMPs such as H istatin[114] This link to the SCPP gene cluster, its proline rich nature, small size, and unique resistance to proteases[13] suggest that SCPPPQ1 may have antimicrobial capacity. Our specific objective was to apply molecular and biophysical techniques to define in silico and in vitro the impact of rat SCPPPQ1 protein on this aggressive bacterium implicated in PD
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