Immunochemical Identification of Immunogenic Proteins from Porphyromonas gingivalis

Authors

Keywords:

Porphyromonas gingivalis, Periodontitis, Antigens, Western Blotting, Immunoglobulin G, Bacterium

Abstract

Porphyromonas gingivalis is a major periodontal pathogen whose immunogenic proteins play essential roles in host-pathogen interactions and represent promising targets for the development of diagnostic and immunotherapeutic strategies. This study aimed to identify immunogenic proteins of Porphyromonas gingivalis through an immunochemical analysis using sera from BALB/c mice immunized with a bacterial crude protein extract (ECPg). The ECPg was prepared from bacterial cultures (strain ATCC 33277) and used as an antigen in an intraperitoneal immunization protocol. The humoral immune response was confirmed by ELISA. Protein composition was analyzed by 12 % SDS-PAGE, whereas immunoreactive proteins were detected by Western blot using immune mouse sera. Fifteen protein bands were identified in the ECPg, of which seven (28, 33, 40, 54, 62, 72, and 104 kDa) were recognized by specific IgG antibodies. The 62-kDa protein (p62) exhibited the strongest immunoreactivity, suggesting that it represents the dominant immunogen within the extract. In silico analysis enabled the putative identification of six immunoreactive proteins as Omp28, Mfa5 fimbrial protein, Omp40, Mfa1 fimbrial protein, phosphotransacetylase, and K gingipain, whereas p62 remained unidentified. These findings demonstrate that the ECPg contains multiple antigens capable of eliciting a specific humoral immune response. Although protein identities remain putative and rely on bioinformatic correlation, this study expands the current knowledge of the antigenic repertoire of Porphyromonas gingivalis and provides a foundation for future structural, functional, and immunological characterization, particularly of the highly immunoreactive p62 protein.

Downloads

Download data is not yet available.

Author Biographies

  • Beltrán-López E., Posgrado en Ciencias Biomédicas, Facultad de Ciencias Químico Biológicas, Universidad Autónoma de Sinaloa

    Evangelina Beltrán López is a Professor/Researcher at the Faculty of Chemical-Biological Sciences of the Autonomous University of Sinaloa (UAS). She holds a Master’s Degree in Forensic Sciences from Universidad Marista de Guadalajara. She is a member of SSIT as an Assistant Researcher. She has published 7 articles in indexed journals and 4 articles in peer-reviewed journals, with 41 citations and an h-index of 4 to date. She has participated in 21 national and international conferences. Throughout her academic career, she has supervised 2 undergraduate theses and 1 master’s thesis in the fields of Biology and Chemistry. Additionally, she serves as a lecturer in the Pharmaceutical Biologist Chemistry (QFB) program, as a thesis advisor/supervisor for undergraduate and master’s students in Genomic Biotechnology, and is currently pursuing a Ph.D. in Biomedical Sciences. Contact e-mail: evabeltran.fcqb@ms.uas.edu.mx; evabeltran@uas.edu.mx

  • Tiznado-Parra N.N., Laboratorio de Biomedicina Molecular, del Posgrado en Ciencias Biomédicas de la Universidad Autónoma de Sinaloa

    She is a graduate student in the Biomedical Sciences Graduate Program at the Faculty of Chemical-Biological Sciences. She holds a Bachelor’s Degree in Genomic Biotechnology from the Autonomous University of Sinaloa, Faculty of Chemical-Biological Sciences. Contact e-mail: nanziuannapaketzallitiznado.fcqb@ms.uas.edu.mx

       
  • Luque-Ortega F., Facultad de Odontología de la Universidad Autónoma de Sinaloa

    He is a Professor/Researcher at the Faculty of Dentistry and holds a Ph.D. in Health Biotechnology from the Autonomous University of Sinaloa. He is a member of the National System of Researchers and Research Fellows (SNII), Level 1, a member of SSIT as an Honorary Researcher, and holds PRODEP recognition. He has published 16 articles in indexed journals and 14 articles in peer-reviewed journals, with 115 citations and an h-index of 7 to date. He has participated in 138 national and international conferences. Throughout his academic career, he has supervised 9 undergraduate theses and 2 master’s theses in the field of Health Sciences. Contact e-mail: fredluque@uas.edu.mx

  • Arámbula-Meraz E., Posgrado en Ciencias Biomédicas de la Universidad Autónoma de Sinaloa

    She is a Professor/Researcher at the Faculty of Chemical-Biological Sciences and holds a Ph.D. in Human Genetics from the Western Biomedical Research Center. She is a Level 1 member of the National System of Researchers (SNII), a member of SSIT as an Honorary Researcher, and holds PRODEP recognition. She has published 41 articles in indexed journals and 39 articles in peer-reviewed journals, with 608 citations and an h-index of 15 to date. She has participated in 220 national and international conferences. Throughout her academic career, she has supervised 30 undergraduate theses, 23 master’s theses, and 7 doctoral theses in the fields of Medicine and Health Sciences. Contact e-mail: eliakymarambula@uas.edu.mx

  • Pineda-Hidalgo K.V., Posgrado en Ciencia y Tecnología de Alimentos, de la Universidad Autónoma de Sinaloa

    She is a Full-Time Research Professor affiliated to Graduate Program in Food Science and Technology at the Autonomous University of Sinaloa, and a Level I member of the National System of Researchers (Sistema Nacional de Investigadoras e Investigadores)

  • Vega-Castillo U., Laboratorio de Biomedicina Molecular del Posgrado en Ciencias Biomédicas, de la Universidad Autónoma de Sinaloa

    He is a student at the Autonomous University of Sinaloa and holds a Master’s Degree in Biomedical Sciences from the Autonomous University of Sinaloa. He has published 1 article in an indexed journal and 2 articles in peer-reviewed journals. He has participated in 6 national and international conferences. Throughout his academic career, he has supervised 1 undergraduate thesis in the field of Microbiology and Health Sciences. He is currently pursuing a Ph.D. in Biomedical Sciences at the Autonomous University of Sinaloa. Contact e-mail: ulisesvega.fcqb@uas.edu.mx

  • López Moreno H.S., Laboratorio de Biomedicina Molecular, Posgrado en Ciencias Biomédicas, Universidad Autónoma de Sinaloa

    He is a Professor/Researcher at the Molecular Biomedicine Laboratory, Faculty of Chemical-Biological Sciences, Autonomous University of Sinaloa, and holds a Ph.D. in Biomedical Sciences awarded by UNAM. He is a Level 2 member of the SNII, an Honorary Researcher of the SSIT, and holds a PRODEP Profile. He has published 45 articles in indexed journals and 4 in peer-reviewed journals, with a total of 398 citations to date and an h-index of 10. He has participated in 62 national and international conferences. Throughout his career, he has supervised 44 undergraduate theses, 18 master's theses, and 5 doctoral dissertations in the fields of Biology and Chemistry. Additionally, he works on Immunobiotechnology and Apoptosis in Giardia lamblia. Contact email: hslmoreno@ms.uas.edu.mx

References

Abramson, J., Adler, J., Dunger, J., Evans, R., Green, T., Pritzel, A., Ronneberger, O., Willmore, L., Ballard, A. J., Bambrick, J., Bodenstein, S. W., Evans, D. A., Hung, C. C., O’Neill, M., Reiman, D., Tunyasuvunakool, K., Wu, Z., Žemgulytė, A., Arvaniti, E., … Jumper, J. M. (2024). Accurate structure prediction of biomolecular interactions with AlphaFold 3. Nature, 630(8016), 493–500. https://doi.org/10.1038/s41586-024-07487-w

Cancro, M. P., & Tomayko, M. M. (2021). Memory B cells and plasma cells: The differentiative continuum of humoral immunity. In Immunological Reviews. John Wiley and Sons Inc. 303 (1), 72–82. https://doi.org/10.1111/imr.13016

Chen, C.-K. C., Denardin, A., Dyer, D. W., Genco, R. J., & Neiders3, M. E. (1991). Human Immunoglobulin G Antibody Response to Iron-Repressible and Other Membrane Proteins of Porphyromonas (Bacteroides) gingivalis. Infection and Immunity. 59 (7), 2427-2433. https://doi.org/10.1128/iai.59.7.2427-2433.1991

Diehl, K.-H., Hull, R., Morton, D., Pfister, R., Rabemampianina, Y., Smith, D., Vidal, J.-M., & Van De Vorstenbosch, C. (2001). A Good Practice Guide to the Administration of Substances and Removal of Blood, Including Routes and Volumes. Journal Of Applied Toxicology 21: 15-23. https://doi.org/10.1002/jat.727

Franca, M., Moura-Costa, L., Meyer, R. J., Trindade, S. C., Da, U., Tunes, R., & Freire, S. M. (2007). Humoral immune response to antigens of Porphyromonas gingivalis ATCC 33277 in chronic periodontitis. Journal of Applied Oral Science. 15 (3), 213-219. https://doi.org/10.1590/S1678-77572007000300011

González, P., Beltrán-López, E., & López-Moreno H.S. (2024). Cloning and recombinant expression of the Immunoglobulin-like variable domains of humans PD-1 and PD-L1 immunoreceptors. QUIBIOUAS Revista De Ciencias Químico Biológicas, 2,1-10. https://revistas.uas.edu.mx/index.php/QBU/article/view/598

Guevara, T., Rodríguez-Banqueri, A., Lasica, A. M., Ksiazek, M., Potempa, B. A., Potempa, J., & Gomis-Rüth, F. X. (2019). Structural determinants of inhibition of Porphyromonas gingivalis gingipain K by KYT-36, a potent, selective, and bioavailable peptidase inhibitor. Scientific Reports, 9(1). https://doi.org/10.1038/s41598-019-41354-3

Hall, M., Hasegawa, Y., Yoshimura, F., & Persson, K. (2018). Structural and functional characterization of shaft, anchor, and tip proteins of the Mfa1 fimbria from the periodontal pathogen Porphyromonas gingivalis. Scientific Reports, 8(1). https://doi.org/10.1038/s41598-018-20067-z

Heidler, T. V., Ernits, K., Ziolkowska, A., Claesson, R., & Persson, K. (2021). Porphyromonas gingivalis fimbrial protein Mfa5 contains a von Willebrand factor domain and an intramolecular isopeptide. Communications Biology, 4(1). https://doi.org/10.1038/s42003-020-01621-w

Hirai, K., Yamaguchi-Tomikawa, T., Eguchi, T., Maeda, H., & Takashiba, S. (2020). Identification and Modification of Porphyromonas gingivalis Cysteine Protease, Gingipain, Ideal for Screening Periodontitis. Frontiers in Immunology, 11. https://doi.org/10.3389/fimmu.2020.01017

Hiratsuka, K., Abiko, Y., Hayakawa, M., Ito, T., Sasahara, H., & Takiguchi, H. (1992). Role of Porphyromonas gingivalis 40-kDa Outer Membrane Protein in the aggregation of P. gingivalis vesicles and Actinomyces viscosus. Archives of oral biology, 37(9), 717–724. https://doi.org/10.1016/0003-9969(92)90078-m

López-Moreno, H S., Correa, D., Laclette, J.P., & Ortiz-Navarrete, V.F. (2003). Identification of CD4 + T cell epitopes of Taenia solium paramyosin. In Parasite Immunology. 25(10), 513-516. https://doi.org/10.1111/j.1365-3024.2003.00658.x

Lunar, I., & Cascales, E. (2021). Molecular Strategies Underlying Porphyromonas gingivalis Virulence. In Journal of Molecular Biology. Academic Press. 433 (7). 166836. https://doi.org/10.1016/j.jmb.2021.166836

Mann, M., Hendrickson, R. C., & Pandey, A. (2001). Analysis of Proteins and Proteomes By Mass Spectrometry. Annual Review of Biochemistry. 70:437-473. https://doi.org/10.1146/annurev.biochem.70.1.437

Messina, B. M., Grippaudo, C., Polizzi, A., Blasi, A., & Isola, G. (2025). The Key Role of Porphyromonas gingivalis in the Pathogenesis of Periodontitis Linked with Systemic Diseases. In Applied Sciences (Switzerland) 15(12). 6847. https://doi.org/10.3390/app15126847

Mysak, J., Podzimek, S., Sommerova, P., Lyuya-Mi, Y., Bartova, J., Janatova, T., Prochazkova, J., & Duskova, J. (2014). Porphyromonas gingivalis: Major periodontopathic pathogen overview. Journal of Immunology Research. Hindawi Publishing Corporation. 476068, https://doi.org/10.1155/2014/476068

Namikoshi, J., Otake, S., Maeba, S., Hayakawa, M., Abiko, Y., & Yamamoto, M. (2003). Specific antibodies induced by nasally administered 40-kDa outer membrane protein of Porphyromonas gingivalis inhibits coaggregation activity of P. gingivalis. Vaccine, 22(2), 250–256. https://doi.org/10.1016/S0264-410X(03)00576-0

Nazir, M., Al-Ansari, A., Al-Khalifa, K., Alhareky, M., Gaffar, B., & Almas, K. (2020). Global Prevalence of Periodontal Disease and Lack of Its Surveillance. Scientific World Journal. 2146160. https://doi.org/10.1155/2020/2146160

Nobre dos Santos-Lima, E. K., Araújo Paiva Andrade Cardoso, K., Mares de Miranda, P., Cirino de Carvalho-Filho, P., Passos Rocha, T., Ferreira de Moura-Costa, L., Olczak, T., Miranda Lopes Falcão, M., Gomes-Filho, I. S., Meyer, R., Tosta Xavier, M., & Castro Trindade, S. (2020). Novel synthetic peptide derived from Porphyromonas gingivalis Lys-gingipain detects IgG-mediated host response in periodontitis. Anaerobe, 61. 102140. https://doi.org/10.1016/j.anaerobe.2019.102140

Oliva, L. E. R., Najera, R. I. S., Zavala, C. E. del P. S. M., Ancona, P. E. L., Elizondo, J. E., Rangel, S. S., Regalado, S. E. F., & Soto, J. M. S. (2022). Porphyromonas: A dental approach. International Journal of Applied Dental Sciences, 8(2), 213–216. https://doi.org/10.22271/oral.2022.v8.i2d.1510

Piedra-Quintero, Z. L., Apodaca-Medina, A. I., Beltrán-López, E., León-Sicairos, C. R., Chávez-Ontiveros, J., Rendón-Maldonado, J. G., Ortiz-Navarrete, V. F., & López-Moreno, H. S. (2015). Immunoproteomic Identification of p29 Antigen as the Elongation Factor-1α of Leishmania mexicana. Vector-Borne and Zoonotic Diseases, 15(7), 449–452. https://doi.org/10.1089/vbz.2014.1712

Reyes, L. (2021). Porphyromonas gingivalis. In Trends in Microbiology (Vol. 29, Number 4, pp. 376–377). Elsevier Ltd. https://doi.org/10.1016/j.tim.2021.01.010

Sanchez, A. A., Moyeda, A. L. G., Ibarra, K. I. J., Pascual, J. B., Rodriguez, D. L., Salazar, V. I. Q., Villarreal, C. A. A., & Soto, J. M. S. (2022). Porphyromonas gingivalis, an Orthodontic point of view. International Journal of Applied Dental Sciences, 8(4), 82–87. https://doi.org/10.22271/oral.2022.v8.i4b.1629

Siemińska, K., Cierpisz, P., Śmiga, M., & Olczak, T. (2021). Porphyromonas gingivalis hmuy and bacteroides vulgatus bvu—a novel competitive heme acquisition strategy. International Journal of Molecular Sciences, 22(5), 1–16. https://doi.org/10.3390/ijms22052237

Slakeski, N., Margetts, M., Moore, C., Czajkowski, L., Barr, I. G., & Reynolds, E. C. (2002). Characterization and expression of a novel Porphyromonas gingivalis outer membrane protein, Omp28. Oral Microbiology Immunology, 17, 150–156., 105–115. https://doi.org/10.1042/0264-6021:3630105

Villa-Medina, M. C., Díaz-Gaxiola, C., Rosales-Reyes, R., Durán-Pérez, S. A., Vega-Castillo, U., Rodríguez-Rochín, J. A., León-Sicairos, C. del R., Beltrán-López, E., & López-Moreno, H. S. (2025). Cloning and Recombinant Expression of the Caspase‐Activated DNase Orthologous Gene of Giardia lamblia. BioMed Research International, 3420875. https://doi.org/10.1155/bmri/3420875

Xu, W., Zhou, W., Wang, H., & Liang, S. (2020). Roles of Porphyromonas gingivalis and its virulence factors in periodontitis. Advances in Protein Chemistry and Structural Biology, 120, 45–84. https://doi.org/10.1016/bs.apcsb.2019.12.001

Yoshida, Y., Sato, M., Nonaka, T., Hasegawa, Y., & Kezuka, Y. (2019). Characterization of the phosphotransacetylase-acetate kinase pathway for ATP production in Porphyromonas gingivalis. Journal of Oral Microbiology, 11(1). https://doi.org/10.1080/20002297.2019.1588086

Beltrán 2026

Downloads

Published

2026-08-19

Data Availability Statement

Graphical Abstract

Issue

No. 5 (2026): QUIBIOUAS Revista de Ciencias Químico Biológicas

Section

Scientific articles

Categories

License

Copyright (c) 2026 QUIBIOUAS Revista de Ciencias Químico Biológicas

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

How to Cite

Beltrán-López, E., Tiznado-Parra, N. N., Luque-Ortega, F., Arámbula-Meraz, E., Vega-Castillo, U., & López Moreno , H. S. . (2026). Immunochemical Identification of Immunogenic Proteins from Porphyromonas gingivalis (K. V. Pineda-Hidalgo , Trans.). QUIBIOUAS Revista De Ciencias Químico Biológicas, 5, 41-53. https://revistas.uas.edu.mx/index.php/QBU/article/view/1915