File Download

There are no files associated with this item.

  Links for fulltext
     (May Require Subscription)
  • Find via Find It@HKUL

Conference Paper: Fluorescence- and 2D gel-based bismuth-proteome of Porphyromonas gingivalis

TitleFluorescence- and 2D gel-based bismuth-proteome of Porphyromonas gingivalis
Authors
Issue Date2016
PublisherSage Publications, Inc. The Journal's web site is located at http://jdr.sagepub.com/
Citation
The 94th General Session & Exhibition of the IADR, 3rd Meeting of the IADR Asia Pacific Region & 35th Annual Meeting of the IADR Korean Division, Seoul, Korea, 22-25 June 2016. In Journal of Dental Research, 2016, v. 95 Spec. Iss. B, abstract no. 95 How to Cite?
AbstractOBJECTIVES: Our recent work shows that the growth of the ‘keystone’ periodontal pathogen Porphyromonas gingivalis could be inhibited by bismuth-based anti-Helicobacter pylori drugs. This study aimed to identify bismuth-associating proteins within P. gingivalis using an intracellular fluorescent probe (Bi3+-TRACER), and explore the underlying molecular mechanisms of bismuth-based inhibition of the bacterium. METHODS: Bi3+-TRACER was prepared by mixing bismuth citrate with TRACER at 1:1 molar ratio. P. gingivalis (ATCC 33277) was cultured in liquid medium at 37°C anaerobically for three days. Bacteria were washed three times and concentrated by PBS, and then added 50 μM of final concentration of Bi3+-TRACER with 30-min incubation. The bacterial suspension was diluted for confocal fluorescence imaging. Bacteria were further irradiated by UV light at 365 nm for 10 min before lysis by sonication, and the labeled bacterial lysates were then subjected to 2D-PAGE gel separation of fluorescence-labeling proteins. The fluorescent spots were imaged, and the gels were further analyzed by silver staining. The corresponding lit-up spots were excised and subjected to peptide mass fingerprinting for protein identification. RESULTS: The Bi3+-TRACER could light up P. gingivalis cells in vivo emitting blue fluorescence with little cytotoxicity. On 2D-PAGE gels, proteins in P. gingivalis cellular lysates were separated based on pI and molecular weights. Multiple protein spots in silver-stained gels were noticed. By comparison, lit-up spots were excised and identified. The identified potential bismuth-associating proteins included Arg- and Lys-gingipains, hemagglutinin, NAD-specific glutamate dehydrongease, Mn/Fe-SOD, thioredoxin and electron transfer flavoprotein subunit α with high fidelity. CONCLUSIONS: The present study suggests that the multiple bismuth-associating proteins identified may be involved in the underlying mechanisms of bismuth-based inhibition of P. gingivalis. Further study is warranted to solidify the details of bismuth action on P. gingivalis for potential clinical applications.
DescriptionOral Session - Microbiology/Immunology-Periodontal Pathogens: no. 95
Persistent Identifierhttp://hdl.handle.net/10722/227492
ISSN
2015 Impact Factor: 4.602
2015 SCImago Journal Rankings: 1.714

 

DC FieldValueLanguage
dc.contributor.authorCheng, T-
dc.contributor.authorSun, H-
dc.contributor.authorLai, YT-
dc.contributor.authorYang, Y-
dc.contributor.authorJin, L-
dc.date.accessioned2016-07-18T09:11:02Z-
dc.date.available2016-07-18T09:11:02Z-
dc.date.issued2016-
dc.identifier.citationThe 94th General Session & Exhibition of the IADR, 3rd Meeting of the IADR Asia Pacific Region & 35th Annual Meeting of the IADR Korean Division, Seoul, Korea, 22-25 June 2016. In Journal of Dental Research, 2016, v. 95 Spec. Iss. B, abstract no. 95-
dc.identifier.issn0022-0345-
dc.identifier.urihttp://hdl.handle.net/10722/227492-
dc.descriptionOral Session - Microbiology/Immunology-Periodontal Pathogens: no. 95-
dc.description.abstractOBJECTIVES: Our recent work shows that the growth of the ‘keystone’ periodontal pathogen Porphyromonas gingivalis could be inhibited by bismuth-based anti-Helicobacter pylori drugs. This study aimed to identify bismuth-associating proteins within P. gingivalis using an intracellular fluorescent probe (Bi3+-TRACER), and explore the underlying molecular mechanisms of bismuth-based inhibition of the bacterium. METHODS: Bi3+-TRACER was prepared by mixing bismuth citrate with TRACER at 1:1 molar ratio. P. gingivalis (ATCC 33277) was cultured in liquid medium at 37°C anaerobically for three days. Bacteria were washed three times and concentrated by PBS, and then added 50 μM of final concentration of Bi3+-TRACER with 30-min incubation. The bacterial suspension was diluted for confocal fluorescence imaging. Bacteria were further irradiated by UV light at 365 nm for 10 min before lysis by sonication, and the labeled bacterial lysates were then subjected to 2D-PAGE gel separation of fluorescence-labeling proteins. The fluorescent spots were imaged, and the gels were further analyzed by silver staining. The corresponding lit-up spots were excised and subjected to peptide mass fingerprinting for protein identification. RESULTS: The Bi3+-TRACER could light up P. gingivalis cells in vivo emitting blue fluorescence with little cytotoxicity. On 2D-PAGE gels, proteins in P. gingivalis cellular lysates were separated based on pI and molecular weights. Multiple protein spots in silver-stained gels were noticed. By comparison, lit-up spots were excised and identified. The identified potential bismuth-associating proteins included Arg- and Lys-gingipains, hemagglutinin, NAD-specific glutamate dehydrongease, Mn/Fe-SOD, thioredoxin and electron transfer flavoprotein subunit α with high fidelity. CONCLUSIONS: The present study suggests that the multiple bismuth-associating proteins identified may be involved in the underlying mechanisms of bismuth-based inhibition of P. gingivalis. Further study is warranted to solidify the details of bismuth action on P. gingivalis for potential clinical applications.-
dc.languageeng-
dc.publisherSage Publications, Inc. The Journal's web site is located at http://jdr.sagepub.com/-
dc.relation.ispartofJournal of Dental Research-
dc.rightsJournal of Dental Research. Copyright © Sage Publications, Inc.-
dc.titleFluorescence- and 2D gel-based bismuth-proteome of Porphyromonas gingivalis-
dc.typeConference_Paper-
dc.identifier.emailCheng, T: chengtfc@hku.hk-
dc.identifier.emailSun, H: hsun@hku.hk-
dc.identifier.emailLai, YT: skytc13@hku.hk-
dc.identifier.emailJin, L: ljjin@hkucc.hku.hk-
dc.identifier.authoritySun, H=rp00777-
dc.identifier.authorityJin, L=rp00028-
dc.identifier.hkuros259722-
dc.identifier.volume95-
dc.identifier.issueSpec. Iss. B-
dc.publisher.placeUnited States-

Export via OAI-PMH Interface in XML Formats


OR


Export to Other Non-XML Formats