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Open Access 01-11-2018 | Original Investigation

Phosphorylcholine is located in Aggregatibacter actinomycetemcomitans fimbrial protein Flp 1

Authors: Riikka Ihalin, Deyu Zhong, Maribasappa Karched, Casey Chen, Sirkka Asikainen

Published in: Medical Microbiology and Immunology | Issue 5-6/2018

Abstract

Phosphorylcholine (ChoP) is covalently incorporated into bacterial surface structures, contributing to host mimicry and promoting adhesion to surfaces. Our aims were to determine the frequency of ChoP display among Aggregatibacter actinomycetemcomitans strains, to clarify which surface structures bear ChoP, and whether ChoP-positivity relates to serum killing. The tested oral (N = 67) and blood isolates (N = 27) represented 6 serotypes. Mab TEPC-15 was used for immunoblotting of cell lysates and fractions and for immunofluorescence microscopy of cell surface-bound ChoP. The lysates were denatured with urea for hidden ChoP or treated with proteinase K to test whether it binds to a protein. Three ChoP-positive and two ChoP-negative strains were subjected to serum killing in the presence/absence of CRP and using Ig-depleted serum as complement source. Cell lysates and the first soluble cellular fraction revealed a < 10 kDa band in immunoblots. Among 94 strains, 27 were ChoP positive. No difference was found in the prevalence of ChoP-positive oral (21/67) and blood (6/27) strains. Immunofluorescence microscopy corresponded to the immunoblot results. Proteinase K abolished ChoP reactivity, whereas urea did not change the negative result. The TEPC-15-reactive protein was undetectable in Δflp1 mutant strain. The survival rate of serotype-b strains in serum was 100% irrespective of ChoP, but that of serotype-a was higher in ChoP-positive (85%) than ChoP-negative (71%) strains. The results suggest that a third of rough-colony strains harbor ChoP and that ChoP is attached to fimbrial subunit protein Flp1. It further seems that ChoP-positivity does not enhance but may reduce A. actinomycetemcomitans susceptibility to serum killing.

Zeisel SH, da Costa KA (2009) Choline: an essential nutrient for public health. Nutr Rev 67:615–623. https://doi.org/10.1111/j.1753-4887.2009.00246.x CrossRefPubMedPubMedCentral

Lysenko E, Richards JC, Cox AD, Stewart A, Martin A, Kapoor M, Weiser JN (2000) The position of phosphorylcholine on the lipopolysaccharide of Haemophilus influenzae affects binding and sensitivity to C-reactive protein-mediated killing. Mol Microbiol 35:234–245CrossRef

Weiser JN, Goldberg JB, Pan N, Wilson L, Virji M (1998) The phosphorylcholine epitope undergoes phase variation on a 43-kilodalton protein in Pseudomonas aeruginosa and on pili of Neisseria meningitidis and Neisseria gonorrhoeae. Infect Immun 66:4263–4267PubMedPubMedCentral

Clark SE, Weiser JN (2013) Microbial modulation of host immunity with the small molecule phosphorylcholine. Infect Immun 81:392–401. https://doi.org/10.1128/IAI.01168-12 CrossRefPubMedPubMedCentral

Cundell DR, Gerard NP, Gerard C, Idanpaan-Heikkila I, Tuomanen EI (1995) Streptococcus pneumoniae anchor to activated human cells by the receptor for platelet-activating factor. Nature 377:435–438. https://doi.org/10.1038/377435a0 CrossRefPubMed

Schenkein HA, Barbour SE, Berry CR, Kipps B, Tew JG (2000) Invasion of human vascular endothelial cells by Actinobacillus actinomycetemcomitans via the receptor for platelet-activating factor. Infect Immun 68:5416–5419CrossRef

Paturel L, Casalta JP, Habib G, Nezri M, Raoult D (2004) Actinobacillus actinomycetemcomitans endocarditis. Clin Microbiol Infect 10:98–118CrossRef

Rahamat-Langendoen JC, van Vonderen MG, Engstrom LJ, Manson WL, van Winkelhoff AJ, Mooi-Kokenberg EA (2011) Brain abscess associated with Aggregatibacter actinomycetemcomitans: case report and review of literature. J Clin Periodontol 38:702–706. https://doi.org/10.1111/j.1600-051X.2011.01737.x CrossRefPubMed

Paju S, Carlson P, Jousimies-Somer H, Asikainen S (2000) Heterogeneity of Actinobacillus actinomycetemcomitans strains in various human infections and relationships between serotype, genotype, and antimicrobial susceptibility. J Clin Microbiol 38:79–84PubMedPubMedCentral

10.

Gmur R, Thurnheer T, Guggenheim B (1999) Dominant cross-reactive antibodies generated during the response to a variety of oral bacterial species detect phosphorylcholine. J Dent Res 78:77–85. https://doi.org/10.1177/00220345990780011201 CrossRefPubMed

11.

Harnett W, Rzepecka J, Houston KM (2010) How do nematodes transfer phosphorylcholine to carbohydrates? Trends Parasitol 26:114–118. https://doi.org/10.1016/j.pt.2009.12.003 CrossRefPubMed

12.

Serino L, Virji M (2000) Phosphorylcholine decoration of lipopolysaccharide differentiates commensal Neisseriae from pathogenic strains: identification of licA-type genes in commensal Neisseriae. Mol Microbiol 35:1550–1559CrossRef

13.

Hegge FT, Hitchen PG, Aas FE, Kristiansen H, Lovold C, Egge-Jacobsen W, Panico M, Leong WY, Bull V, Virji M, Morris HR, Dell A, Koomey M (2004) Unique modifications with phosphocholine and phosphoethanolamine define alternate antigenic forms of Neisseria gonorrhoeae type IV pili. Proc Natl Acad Sci USA 101:10798–10803. https://doi.org/10.1073/pnas.0402397101 CrossRefPubMed

14.

Clark SE, Snow J, Li J, Zola TA, Weiser JN (2012) Phosphorylcholine allows for evasion of bactericidal antibody by Haemophilus influenzae. PLoS Pathog 8:e1002521. https://doi.org/10.1371/journal.ppat.1002521 CrossRefPubMedPubMedCentral

15.

Volanakis JE (2001) Human C-reactive protein: expression, structure, and function. Mol Immunol 38:189–197CrossRef

16.

Sundqvist G, Johansson E (1982) Bactericidal effect of pooled human serum on Bacteroides melaninogenicus, Bacteroides asaccharolyticus and Actinobacillus actinomycetemcomitans. Scand J Dent Res 90:29–36PubMed

17.

Asakawa R, Komatsuzawa H, Kawai T, Yamada S, Goncalves RB, Izumi S, Fujiwara T, Nakano Y, Suzuki N, Uchida Y, Ouhara K, Shiba H, Taubman MA, Kurihara H, Sugai M (2003) Outer membrane protein 100, a versatile virulence factor of Actinobacillus actinomycetemcomitans. Mol Microbiol 50:1125–1139CrossRef

18.

Asikainen S, Lai CH, Alaluusua S, Slots J (1991) Distribution of Actinobacillus actinomycetemcomitans serotypes in periodontal health and disease. Oral Microbiol Immunol 6:115–118CrossRef

19.

Dogan B, Saarela MH, Jousimies-Somer H, Alaluusua S, Asikainen S (1999) Actinobacillus actinomycetemcomitans serotype e–biotypes, genetic diversity and distribution in relation to periodontal status. Oral Microbiol Immunol 14:98–103CrossRef

20.

Wang Y, Chen C (2005) Mutation analysis of the flp operon in Actinobacillus actinomycetemcomitans. Gene 351:61–71. https://doi.org/10.1016/j.gene.2005.02.010 CrossRefPubMed

21.

Karched M, Paul-Satyaseela M, Asikainen S (2007) A simple viability-maintaining method produces homogenic cell suspensions of autoaggregating wild-type Actinobacillus actinomycetemcomitans. J Microbiol Methods 68:46–51. https://doi.org/10.1016/j.mimet.2006.06.004 CrossRefPubMed

22.

Asikainen S, Chen C, Slots J (1995) Actinobacillus actinomycetemcomitans genotypes in relation to serotypes and periodontal status. Oral Microbiol Immunol 10:65–68CrossRef

23.

Kittichotirat W, Bumgarner RE, Asikainen S, Chen C (2011) Identification of the pangenome and its components in 14 distinct Aggregatibacter actinomycetemcomitans strains by comparative genomic analysis. PLoS One 6:e22420. https://doi.org/10.1371/journal.pone.0022420 CrossRefPubMedPubMedCentral

24.

Saarela M, Asikainen S, Alaluusua S, Pyhälä L, Lai CH, Jousimies-Somer H (1992) Frequency and stability of mono- or poly-infection by Actinobacillus actinomycetemcomitans serotypes a, b, c, d or e. Oral Microbiol Immunol 7:277–279CrossRef

25.

Dogan B, Antinheimo J, Cetiner D, Bodur A, Emingil G, Buduneli E, Uygur C, Firatli E, Lakio L, Asikainen S (2003) Subgingival microflora in Turkish patients with periodontitis. J Periodontol 74:803–814CrossRef

26.

Paul-Satyaseela M, Karched M, Bian Z, Ihalin R, Boren T, Arnqvist A, Chen C, Asikainen S (2006) Immunoproteomics of Actinobacillus actinomycetemcomitans outer-membrane proteins reveal a highly immunoreactive peptidoglycan-associated lipoprotein. J Med Microbiol 55:931–942. https://doi.org/10.1099/jmm.0.46470-0 CrossRefPubMed

27.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

28.

Wilson ME (1991) The heat-modifiable outer membrane protein of Actinobacillus actinomycetemcomitans: relationship to OmpA proteins. Infect Immun 59:2505–2507PubMedPubMedCentral

29.

Serino L, Virji M (2002) Genetic and functional analysis of the phosphorylcholine moiety of commensal Neisseria lipopolysaccharide. Mol Microbiol 43:437–448CrossRef

30.

Wang Y, Liu A, Chen C (2005) Genetic basis for conversion of rough-to-smooth colony morphology in Actinobacillus actinomycetemcomitans. Infect Immun 73:3749–3753. https://doi.org/10.1128/IAI.73.6.3749-3753.2005 CrossRefPubMedPubMedCentral

31.

Inoue T, Tanimoto I, Ohta H, Kato K, Murayama Y, Fukui K (1998) Molecular characterization of low-molecular-weight component protein, Flp, in Actinobacillus actinomycetemcomitans fimbriae. Microbiol Immunol 42:253–258CrossRef

32.

Kachlany SC, Planet PJ, Desalle R, Fine DH, Figurski DH, Kaplan JB (2001) flp-1, the first representative of a new pilin gene subfamily, is required for non-specific adherence of Actinobacillus actinomycetemcomitans. Mol Microbiol 40:542–554CrossRef

33.

Rosan B, Slots J, Lamont RJ, Listgarten MA, Nelson GM (1988) Actinobacillus actinomycetemcomitans fimbriae. Oral Microbiol Immunol 3:58–63CrossRef

34.

Inoue T, Ohta H, Tanimoto I, Shingaki R, Fukui K (2000) Heterogeneous post-translational modification of Actinobacillus actinomycetemcomitans fimbrillin. Microbiol Immunol 44:715–718CrossRef

35.

Tomich M, Fine DH, Figurski DH (2006) The TadV protein of Actinobacillus actinomycetemcomitans is a novel aspartic acid prepilin peptidase required for maturation of the Flp1 pilin and TadE and TadF pseudopilins. J Bacteriol 188:6899–6914CrossRef

36.

Warren MJ, Jennings MP (2003) Identification and characterization of pptA: a gene involved in the phase-variable expression of phosphorylcholine on pili of Neisseria meningitidis. Infect Immun 71:6892–6898CrossRef

37.

Schenkein HA, Gunsolley JC, Best AM, Harrison MT, Hahn CL, Wu J, Tew JG (1999) Antiphosphorylcholine antibody levels are elevated in humans with periodontal diseases. Infect Immun 67:4814–4818PubMedPubMedCentral

38.

Humphries HE, High NJ (2002) The role of licA phase variation in the pathogenesis of invasive disease by Haemophilus influenzae type b. FEMS Immunol Med Microbiol 34:221–230CrossRef

39.

Kittichotirat W, Bumgarner RE, Chen C (2016) Evolutionary Divergence of Aggregatibacter actinomycetemcomitans. J Dent Res 95:94–101. https://doi.org/10.1177/0022034515608163 CrossRefPubMedPubMedCentral

40.

Chen C, Wang T, Chen W (2010) Occurrence of Aggregatibacter actinomycetemcomitans serotypes in subgingival plaque from United States subjects. Mol Oral Microbiol 25:207–214. https://doi.org/10.1111/j.2041-1014.2010.00567.x CrossRefPubMed

41.

Mombelli A, Gmur R, Lang NP, Corbert E, Frey J (1999) Actinobacillus actinomycetemcomitans in Chinese adults. Serotype distribution and analysis of the leukotoxin gene promoter locus. J Clin Periodontol 26:505–510CrossRef

42.

Zambon JJ, Umemoto T, De Nardin E, Nakazawa F, Christersson LA, Genco RJ (1988) Actinobacillus actinomycetemcomitans in the pathogenesis of human periodontal disease. Adv Dent Res 2:269–274. https://doi.org/10.1177/08959374880020021101 CrossRefPubMed

43.

Mattila K, Vesanen M, Valtonen V, Nieminen M, Palosuo T, Rasi V, Asikainen S (2002) Effect of treating periodontitis on C-reactive protein levels: a pilot study. BMC Infect Dis 2:30CrossRef

Title: Phosphorylcholine is located in Aggregatibacter actinomycetemcomitans fimbrial protein Flp 1
Authors: Riikka Ihalin
Deyu Zhong
Maribasappa Karched
Casey Chen
Sirkka Asikainen
Publication date: 01-11-2018
Publisher: Springer Berlin Heidelberg
Published in: Medical Microbiology and Immunology / Issue 5-6/2018
Print ISSN: 0300-8584
Electronic ISSN: 1432-1831
DOI: https://doi.org/10.1007/s00430-018-0554-1

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