Anti-HmuY Antibodies Specifically Recognize Porphyromonas gingivalis HmuY Protein but Not Homologous Proteins in Other Periodontopathogens

Abstract

Given the emerging evidence of an association between periodontal infections and systemic conditions, the search for specific methods to detect the presence of P. gingivalis, a principal etiologic agent in chronic periodontitis, is of high importance. The aim of this study was to characterize antibodies raised against purified P. gingivalis HmuY protein and selected epitopes of the HmuY molecule. Since other periodontopathogens produce homologs of HmuY, we also aimed to characterize responses of antibodies raised against the HmuY protein or its epitopes to the closest homologous proteins from Prevotella intermedia and Tannerella forsythia. Rabbits were immunized with purified HmuY protein or three synthetic, KLH-conjugated peptides, derived from the P. gingivalis HmuY protein. The reactivity of anti-HmuY antibodies with purified proteins or bacteria was determined using Western blotting and ELISA assay. First, we found homologs of P. gingivalis HmuY in P. intermedia (PinO and PinA proteins) and T. forsythia (Tfo protein) and identified corrected nucleotide and amino acid sequences of Tfo. All proteins were overexpressed in E. coli and purified using ion-exchange chromatography, hydrophobic chromatography and gel filtration. We demonstrated that antibodies raised against P. gingivalis HmuY are highly specific to purified HmuY protein and HmuY attached to P. gingivalis cells. No reactivity between P. intermedia and T. forsythia or between purified HmuY homologs from these bacteria and anti-HmuY antibodies was detected. The results obtained in this study demonstrate that P. gingivalis HmuY protein may serve as an antigen for specific determination of serum antibodies raised against this bacterium.

PgRsp Is a Novel Redox-Sensing Transcription Regulator Essential for Porphyromonas gingivalis Virulence

“…Mutant strains were maintained in the presence of 1 µg/mL erythromycin and complemented mutant strains in the presence of 1 µg/mL of tetracycline and 1 µg/mL erythromycin. Prevotella intermedia strain 17 (GenBank: CP003502.1 and CP003503.1) and Tannerella forsythia (ATCC 43037) were maintained as described previously [37]. Streptococcus gordonii (ATCC 10558) was grown in TSB agar or liquid medium (5% TBS) under an increased concentration of CO 2 using the Atmosphere generation system (Thermo Scientific, Waltham, MA, USA) as reported previously [16].…”

ALELOPATIA DE EXTRATOS AQUOSOS DE Eragrostis lugens Nees. NA GERMINAÇÃO E CRESCIMENTO INICIAL DE Oryza sativa L

“…A lipoproteína HmuY é específica de P. gingivalis e anticorpos anti-HmuY não reconhecem proteínas homólogas em outros periodontopatógenos (OLCZAK et al, 2010;ŚMIGA et al, 2015). HmuY apresenta-se associada à membrana externa de P.…”

Role ofPorphyromonas gingivalisHmuY in Immunopathogenesis of Chronic Periodontitis

“…Indeed, we found that anti-HmuY antibodies inhibited P. gingivalis growth and biofilm formation [ 91 ]. Antibodies directed against P. gingivalis A7436 and ATCC 33277 HmuY are highly specific for the purified HmuY protein and cell-bound HmuY and do not cross-react with HmuY homologs found in P. intermedia and T. forsythia [ 102 ]. This suggests that P. gingivalis ATCC 33277, W83, and A7436 HmuY can serve as a specific antigen for serum determination of antibodies raised against this bacterium in patients with chronic periodontitis [ 103 ].…”