Head of the Laboratory

• Prof. dr hab. Teresa Olczak

Staff

• Dr Michał Śmiga

PhD Students

• Mgr inż. Klaudia Siemińska
• Mgr inż. Paulina Ślęzak
• Mgr inż. Patryk Cierpisz

Contact:

Laboratory: room No. 3.39, phone number: +48 71375 2394
Head: room No. 3.25, phone number: +48 71 375 2612
Staff and PhD Students: room No. 3.26, phone number: +48 71 375 2304

Research topics and the most important results

• Mechanisms of iron and heme acquisition in bacteria – periodontal diseases and their relationship with human systemic diseases.

Periodontal diseases belong to a group of multifactorial, infectious diseases, initiated by an ecological shift in the composition of subgingival biofilm and an exaggerated host immune response towards agents produced by bacteria, which results in inflammation and destruction of tooth-supporting tissues. Recent years brought the emerging evidence of an association between periodontal diseases and systemic diseases, such as diabetes, cardiovascular diseases, Alzheimer’s disease. Porphyromonas gingivalis, one of the main etiologic agents and key pathogen responsible for initiation and progression of chronic periodontitis, to survive and initiate infection requires iron and heme, which are acquired by the novel, characterized by our group, Hmu system, with the leading role played by a unique hemophore-like HmuY protein. To increase its own virulence, P. gingivalis may also use mechanisms of heme acquisition utilized by other commensal/pathogenic bacteria, such as Streptococcus gordonii, Tannerella forsythia, Prevotella intermedia, as well as host hemoproteins, which results in dysbiosis in oral microbiome. Moreover, mouth-gut bacterial transmission causes that P. gingivalis invading and colonizing the gut is also able to use heme acquisition systems of Bacteroides vulgatus and Bacteroides fragilis, increasing tendency to cause dysbiosis in the gut microbiome, thus influence the host health through participation in systemic inflammatory diseases.

Techniques and methods used

• Molecular biology techniques (cloning, PCR, RT-PCR, EMSA)
• Bacterial cultures (Escherichia coli, Porphyromonas gingivalis, Tannerella forsythia, Prevotella intermedia, Bacteroides fragilis, Streptococcus gordonii)
• Mammalian cell cultures
• Overexpression and purification of recombinant proteins
• Protein purification (chromatographic and electrophoretic methods, low-pressure chromatography, FPLC, HPLC)
• Mutagenesis
• Spectroscopic methods
• Analyses of interactions: protein-protein, protein-heme and protein-DNA

Example doctoral dissertation topics

1. “Characterization of HmuY lipoprotein from Porphyromonas gingivalis“, Halina Wójtowicz, 2011.
2. “Characterization of proteins encoded on the hmu operon in Porphyromonas gingivalis and other bacteria”, Marcin Bielecki, 2015.
3. “Involvement of PgFur protein in virulence of Porphyromonas gingivalis“, Michał Śmiga, 2018.

Research projects

Current

• NCN OPUS 17 (2019/33/B/NZ6/00292) “Hemophore-like proteins of anaerobic Bacteroidetes – important virulence factors of key pathogens of human microbiome, related with inflammatory chronic diseases” (2020-2023).
  Head of the project: prof. dr hab. Teresa Olczak.

Accomplished

• NCN OPUS 12 (2016/23/B/NZ6/00080): “Synergistic mechanisms of heme acquisition used by periodontopathogens under physiological conditions and in periodontal diseases associated with systemic diseases” (2017-2021).
  Head of the project: prof. dr hab. Teresa Olczak.
• NCN OPUS 9 (2015/17/B/NZ6/01969): “Porphyromonas gingivalis protein and its homologs from other periodontopathogens – a novel group of bacterial virulence factors”(2016-2019).
  Head of the project: prof. dr hab. Teresa Olczak.
• NCN OPUS 8 (2014/15/B/NZ6/01723): “Regulation of mechanisms of iron and heme acquisition and virulence in Porphyromonas gingivalis with involvement of PgFur protein and small RNAs” (2015-2018).
  Head of the project: prof. dr hab. Teresa Olczak.
• NCN PRELUDIUM 8 (2014/15/N/NZ6/01718): “Cooperation of PgFur with CRISPR-associated proteins and selected transcriptional regulatory factors in mechanisms of Porphyromonas gingivalis virulence” (2015-2018).
  Head of the project: dr Michał Śmiga

Laboratory equipment

• Anaerobic Workstation
• Double-beam spectrophotometer
• Fast protein liquid chromatography system
• Real-time PCR thermocycler

Publications

1. Siemińska K., Cierpisz P., Śmiga M., Olczak T. (2021) Porphyromonas gingivalis HmuY and Bacteroides vulgatus Bvu-a novel competitive heme acquisition strategy. Int J Mol Sci. 24;22(5):2237
2. Ślęzak P., Śmiga M., Smalley JW., Siemińska K., Olczak T. (2020) Porphyromonas gingivalis HmuY and Streptococcus gordonii GAPDH-Novel Heme Acquisition Strategy in the Oral Microbiome. Int J Mol Sci 10;21(11):4150
3. Śmiga M., Ślęzak P., Siemińska K., Olczak T. (2020) Mechanizmy wirulencji wykorzystywane w patogenezie chorób przyzębia przez bakterie Porphyromonas gingivalis. Postepy Hig Med Dosw 74: 247-258
4. Bielecki M., Antonyuk S., Strange RW., Siemińska K., Smalley JW., Mackiewicz P., Śmiga M., Cowan M., Capper MJ., Ślęzak P., Olczak M., Olczak T. (2020) Prevotella intermedia produces two proteins homologous to Porphyromonas gingivalis HmuY but with different heme coordination mode. Biochem J. 477(2):381-405
5. Nobre Dos Santos-Lima EK., 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 IS., 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 7(61):102140
6. Śmiga M., Olczak T. (2019) PgRsp Is a Novel Redox-Sensing Transcription Regulator Essential for Porphyromonas gingivalis Virulence. Microorganisms 12:623
7. Śmiga M, Bielecki M, Olczak M, Olczak T. (2019) Porphyromonas gingivalis PgFur Is a Member of a Novel Fur Subfamily With Non-canonical Function. Front Cell Infect Microbiol 9:233.
8. Śmiga M, Stępień P, Olczak M, Olczak T. (2019) PgFur participates differentially in expression of virulence factors in more virulent A7436 and less virulent ATCC 33277 Porphyromonas gingivalis strains. BMC Microbiol 19(1):127.
9. Bielecki M, Antonyuk S, Strange RW, Smalley JW, Mackiewicz P, Śmiga M, Stępień P, Olczak M, Olczak T. (2018) Tannerella forsythia Tfo belongs to Porphyromonas gingivalis HmuY-like family of proteins but differs in heme-binding properties. Biosci Rep 22;38(5).
10. Olczak T, Śmiga M, Kwiecień A, Bielecki M, Wróbel R, Olczak M, Ciunik Z. (2017) Antimicrobial activity of stable hemiaminals against Porphyromonas gingivalis. Anaerobe. 44:27-33.
11. Smalley JW, Olczak T (2017) Haem acquisition mechanisms of Porphyromonas gingivalis – strategies used in polymicrobial community in a haem-limited host environment. Mol Oral Microbiol. 32(1):1-23.
12. Gmiterek A, Kłopot A, Wójtowicz H, Trindade SC, Olczak M, Olczak T (2016) Immune response of macrophages induced by Porphyromonas gingivalis requires HmuY protein. Immunobiology 221(12):1382-1394
13. Carvalho-Filho PC, Gomes-Filho IS, Meyer R, Olczak T, Xavier MT, Trindade SC (2016) Role of Porphyromonas gingivalis HmuY in immunopathogenesis of chronic periodontitis. Mediators Inflamm 2016:7465852
14. Olczak T, Sosicka P, Olczak M (2015) HmuY is an important virulence factor for Porphyromonas gingivalis growth in the heme-limited host environment and infection of macrophages. Biochem Biophys Res Commun, 2015, 467, 748-753.
15. Benedyk M, Byrne DP, Glowczyk I, Potempa J, Olczak M, Olczak T, Smalley JW (2015) Pyocyanin, a contributory factor in haem acquisition and virulence enhancement of Porphyromonas gingivalis in the lung. PloS One 10(2):e0118319
16. Śmiga M, Bielecki M, Olczak M, Smalley JW, Olczak T (2015) Anti-HmuY antibodies specifically recognize Porphyromonas gingivalis HmuY protein but not homologous proteins in other periodontopathogens. PloS One 10(2):e0117508
17. Ciuraszkiewicz J, Śmiga M, Mackiewicz P, Gmiterek A, Bielecki M, Olczak M, Olczak T (2014) Fur homolog regulates Porphyromonas gingivalis virulence under low-iron/heme conditions through a complex regulatory network. Mol Oral Microbiol 29:333-353
18. Antonyuk SV, Olczak M, Olczak T, Ciuraszkiewicz J, Strange RW (2014) The structure of a purple acid phosphatase involved in plant growth and pathogen defence exhibits a novel immunoglobulin-like fold. IUCrJ 1:101-109
19. Gmiterek A, Wójtowicz H, Mackiewicz P, Radwan-Oczko M, Kantorowicz M, Chomyszyn-Gajewska M, Frąszczak M, Bielecki M, Olczak M, Olczak T (2013) The unique hmuY gene sequence as a specific marker of Porphyromonas gingivalis. PLoS One 8(7):e67719
20. Byrne DP, Potempa J, Olczak T, Smalley JW (2013) Evidence of mutualism between two periodontal pathogens: Co-operative haem acquisition by the HmuY haemophore of Porphyromonas gingivalis and the cysteine protease interpain A (InpA) of Prevotella intermedia. Mol Oral Microbiol 28:219-229
21. Wójtowicz H, Bielecki M, Wojaczyński J, Olczak M, Smalley JW, Olczak T (2013) The Porphyromonas gingivalis HmuY haemophore binds gallium(iii), zinc(ii), cobalt(iii), manganese(iii), nickel(ii), and copper(ii) protoporphyrin IX but in a manner different to iron(iii) protoporphyrin IX. Metallomics 5:343-351
22. Wojaczyński J, Wójtowicz H, Bielecki M, Olczak M, Smalley JW, Latos-Grażyński L, Olczak T (2011) Iron(III) mesoporphyrin IX and iron(III) deuteroporphyrin IX bind to the Porphyromonas gingivalis HmuY hemophore. Biochem Biophys Res Commun 411:299-304
23. Smalley JW, Byrne DP, Birss AJ, Wojtowicz H, Sroka A, Potempa J, Olczak T (2011) HmuY haemophore and gingipain proteases constitute a unique syntrophic system of haem acquisition by Porphyromonas gingivalis. PLoS One 6(2):e17182
24. Wójtowicz H, Guevara T, Tallant C, Olczak M, Sroka A, Potempa J, Solà M, Olczak T, Gomis-Rüth FX (2009) Unique structure and stability of HmuY, a novel heme-binding protein of Porphyromonas gingivalis. PLoS Pathogens 5(5):e1000419