In Porphyromonas gingivalis VimF Is Involved in Gingipain Maturation through the Transfer of Galactose
Previously, we have reported that gingipain activity in Porphyromonas gingivalis, the major causative agent in adult periodontitis, is post-translationally regulated by the unique Vim proteins including VimF, a putative glycosyltransferase. To further characterize VimF, an isogenic mutant defective...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Public Library of Science
2013
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3663753/ https://www.ncbi.nlm.nih.gov/pubmed/23717416 http://dx.doi.org/10.1371/journal.pone.0063367 |
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author | Muthiah, Arun S. Aruni, Wilson Robles, Antonette G. Dou, Yuetan Roy, Francis Fletcher, Hansel M. |
author_facet | Muthiah, Arun S. Aruni, Wilson Robles, Antonette G. Dou, Yuetan Roy, Francis Fletcher, Hansel M. |
author_sort | Muthiah, Arun S. |
collection | PubMed |
description | Previously, we have reported that gingipain activity in Porphyromonas gingivalis, the major causative agent in adult periodontitis, is post-translationally regulated by the unique Vim proteins including VimF, a putative glycosyltransferase. To further characterize VimF, an isogenic mutant defective in this gene in a different P. gingivalis genetic background was evaluated. In addition, the recombinant VimF protein was used to further confirm its glycosyltransferase function. The vimF-defective mutant (FLL476) in the P. gingivalis ATCC 33277 genetic background showed a phenotype similar to that of the vimF-defective mutant (FLL95) in the P. gingivalis W83 genetic background. While hemagglutination was not detected and autoaggregation was reduced, biofilm formation was increased in FLL476. HeLa cells incubated with P. gingivalis FLL95 and FLL476 showed a 45% decrease in their invasive capacity. Antibodies raised against the recombinant VimF protein in E. coli immunoreacted only with the deglycosylated native VimF protein from P. gingivalis. In vitro glycosyltransferase activity for rVimF was observed using UDP-galactose and N-acetylglucosamine as donor and acceptor substrates, respectively. In the presence of rVimF and UDP-galactose, a 60 kDa protein from the extracellular fraction of FLL95 which was identified by mass spectrometry as Rgp gingipain, immunoreacted with the glycan specific mAb 1B5 antibody. Taken together, these results suggest the VimF glycoprotein is a galactosyltransferase that may be specific for gingipain glycosylation. Moreover, galatose is vital for the growing glycan chain. |
format | Online Article Text |
id | pubmed-3663753 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-36637532013-05-28 In Porphyromonas gingivalis VimF Is Involved in Gingipain Maturation through the Transfer of Galactose Muthiah, Arun S. Aruni, Wilson Robles, Antonette G. Dou, Yuetan Roy, Francis Fletcher, Hansel M. PLoS One Research Article Previously, we have reported that gingipain activity in Porphyromonas gingivalis, the major causative agent in adult periodontitis, is post-translationally regulated by the unique Vim proteins including VimF, a putative glycosyltransferase. To further characterize VimF, an isogenic mutant defective in this gene in a different P. gingivalis genetic background was evaluated. In addition, the recombinant VimF protein was used to further confirm its glycosyltransferase function. The vimF-defective mutant (FLL476) in the P. gingivalis ATCC 33277 genetic background showed a phenotype similar to that of the vimF-defective mutant (FLL95) in the P. gingivalis W83 genetic background. While hemagglutination was not detected and autoaggregation was reduced, biofilm formation was increased in FLL476. HeLa cells incubated with P. gingivalis FLL95 and FLL476 showed a 45% decrease in their invasive capacity. Antibodies raised against the recombinant VimF protein in E. coli immunoreacted only with the deglycosylated native VimF protein from P. gingivalis. In vitro glycosyltransferase activity for rVimF was observed using UDP-galactose and N-acetylglucosamine as donor and acceptor substrates, respectively. In the presence of rVimF and UDP-galactose, a 60 kDa protein from the extracellular fraction of FLL95 which was identified by mass spectrometry as Rgp gingipain, immunoreacted with the glycan specific mAb 1B5 antibody. Taken together, these results suggest the VimF glycoprotein is a galactosyltransferase that may be specific for gingipain glycosylation. Moreover, galatose is vital for the growing glycan chain. Public Library of Science 2013-05-24 /pmc/articles/PMC3663753/ /pubmed/23717416 http://dx.doi.org/10.1371/journal.pone.0063367 Text en © 2013 Muthiah et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Muthiah, Arun S. Aruni, Wilson Robles, Antonette G. Dou, Yuetan Roy, Francis Fletcher, Hansel M. In Porphyromonas gingivalis VimF Is Involved in Gingipain Maturation through the Transfer of Galactose |
title | In Porphyromonas gingivalis VimF Is Involved in Gingipain Maturation through the Transfer of Galactose |
title_full | In Porphyromonas gingivalis VimF Is Involved in Gingipain Maturation through the Transfer of Galactose |
title_fullStr | In Porphyromonas gingivalis VimF Is Involved in Gingipain Maturation through the Transfer of Galactose |
title_full_unstemmed | In Porphyromonas gingivalis VimF Is Involved in Gingipain Maturation through the Transfer of Galactose |
title_short | In Porphyromonas gingivalis VimF Is Involved in Gingipain Maturation through the Transfer of Galactose |
title_sort | in porphyromonas gingivalis vimf is involved in gingipain maturation through the transfer of galactose |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3663753/ https://www.ncbi.nlm.nih.gov/pubmed/23717416 http://dx.doi.org/10.1371/journal.pone.0063367 |
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