Europe PMC

This website requires cookies, and the limited processing of your personal data in order to function. By using the site you are agreeing to this as outlined in our privacy notice and cookie policy.

Binding of vitronectin to opa-expressing Neisseria gonorrhoeae mediates invasion of HeLa cells.

Author information

Affiliations

  • 1. Abteilung Infektionsbiologie, Max-Planck-Institut für Biologie, Tübingen, Germany.
      Authors
    • Gómez-Duarte OG 1
    (1 author)

ORCIDs linked to this article

Infection and Immunity, 01 Sep 1997, 65(9):3857-3866
https://doi.org/10.1128/iai.65.9.3857-3866.1997 PMID: 9284164 PMCID: PMC175551

Free full text in Europe PMC

Abstract 

Neisseria gonorrhoeae induces local infections in the human genitourinary tract and can disseminate to other organs to cause severe disease. Blood-derived factors present in the genital mucosa have been suggested to facilitate the spread of N. gonorrhoeae in disseminated gonococcal infections. Using gentamicin invasion assays and confocal microscopy, we observed a strong stimulatory effect of fetal calf serum (FCS) on the gonococcal invasion of HeLa cells. FCS-mediated invasion was dependent on the expression of the epithelial cell invasion-associated Opa protein (plasmid-encoded Opa50 or its chromosomal homolog Opa30), while N. gonorrhoeae expressing noninvasive Opa proteins (Opa(51-60)) or no Opa protein (Opa-) was not invasive even in the presence of FCS. Incubation of N. gonorrhoeae MS11 with biotinylated FCS revealed a 78-kDa protein as the prominent protein binding to Opa50- or Opa30-expressing gonococci. This protein was recognized by antibodies against vitronectin (VN) in Western blots. Purified human or bovine VN efficiently bound to Opa50-expressing gonococci, while binding to noninvasive Opa- or Opa52-expressing gonococci was significantly lower. Binding of VN was inhibited by heparin in a concentration-dependent manner, indicating that the heparin binding sites present in VN or Opa50 may play an essential role in this interaction. Based on gentamicin invasion assays and confocal microscopy studies, VN binding was associated with an increased invasion of Opa50- and Opa30-expressing gonococci into HeLa cells. The ability of VN to mediate entry into epithelial cells may constitute an important event in the pathogenesis of local as well as disseminated gonococcal infections.

Free full text 

Logo of iaiLink to Publisher's site
Infect Immun. 1997 Sep; 65(9): 3857–3866.
PMCID: PMC175551
PMID: 9284164

Binding of vitronectin to opa-expressing Neisseria gonorrhoeae mediates invasion of HeLa cells.

O G Gómez-Duarte, M Dehio, C A Guzmán, G S Chhatwal, C Dehio, and T F Meyer
Author information Copyright and License information Disclaimer

Abstract

Neisseria gonorrhoeae induces local infections in the human genitourinary tract and can disseminate to other organs to cause severe disease. Blood-derived factors present in the genital mucosa have been suggested to facilitate the spread of N. gonorrhoeae in disseminated gonococcal infections. Using gentamicin invasion assays and confocal microscopy, we observed a strong stimulatory effect of fetal calf serum (FCS) on the gonococcal invasion of HeLa cells. FCS-mediated invasion was dependent on the expression of the epithelial cell invasion-associated Opa protein (plasmid-encoded Opa50 or its chromosomal homolog Opa30), while N. gonorrhoeae expressing noninvasive Opa proteins (Opa(51-60)) or no Opa protein (Opa-) was not invasive even in the presence of FCS. Incubation of N. gonorrhoeae MS11 with biotinylated FCS revealed a 78-kDa protein as the prominent protein binding to Opa50- or Opa30-expressing gonococci. This protein was recognized by antibodies against vitronectin (VN) in Western blots. Purified human or bovine VN efficiently bound to Opa50-expressing gonococci, while binding to noninvasive Opa- or Opa52-expressing gonococci was significantly lower. Binding of VN was inhibited by heparin in a concentration-dependent manner, indicating that the heparin binding sites present in VN or Opa50 may play an essential role in this interaction. Based on gentamicin invasion assays and confocal microscopy studies, VN binding was associated with an increased invasion of Opa50- and Opa30-expressing gonococci into HeLa cells. The ability of VN to mediate entry into epithelial cells may constitute an important event in the pathogenesis of local as well as disseminated gonococcal infections.

Full Text

The Full Text of this article is available as a PDF (573K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Apicella MA, Ketterer M, Lee FK, Zhou D, Rice PA, Blake MS. The pathogenesis of gonococcal urethritis in men: confocal and immunoelectron microscopic analysis of urethral exudates from men infected with Neisseria gonorrhoeae. J Infect Dis. 1996 Mar;173(3):636–646. [Abstract] [Google Scholar]
  • Arko RJ, Chen CY, Schalla WO, Sarafian SK, Taylor CL, Knapp JS, Morse SA. Binding of S protein by Neisseria gonorrhoeae and potential role in invasion. J Clin Microbiol. 1991 Jan;29(1):70–75. [Europe PMC free article] [Abstract] [Google Scholar]
  • Bessen D, Gotschlich EC. Interactions of gonococci with HeLa cells: attachment, detachment, replication, penetration, and the role of protein II. Infect Immun. 1986 Oct;54(1):154–160. [Europe PMC free article] [Abstract] [Google Scholar]
  • Bhat KS, Gibbs CP, Barrera O, Morrison SG, Jähnig F, Stern A, Kupsch EM, Meyer TF, Swanson J. The opacity proteins of Neisseria gonorrhoeae strain MS11 are encoded by a family of 11 complete genes. Mol Microbiol. 1991 Aug;5(8):1889–1901. [Abstract] [Google Scholar]
  • Bihimaier A, Römling U, Meyer TF, Tümmler B, Gibbs CP. Physical and genetic map of the Neisseria gonorrhoeae strain MS11-N198 chromosome. Mol Microbiol. 1991 Oct;5(10):2529–2539. [Abstract] [Google Scholar]
  • Blake MS, Blake CM, Apicella MA, Mandrell RE. Gonococcal opacity: lectin-like interactions between Opa proteins and lipooligosaccharide. Infect Immun. 1995 Apr;63(4):1434–1439. [Europe PMC free article] [Abstract] [Google Scholar]
  • Chen T, Belland RJ, Wilson J, Swanson J. Adherence of pilus- Opa+ gonococci to epithelial cells in vitro involves heparan sulfate. J Exp Med. 1995 Aug 1;182(2):511–517. [Europe PMC free article] [Abstract] [Google Scholar]
  • Chhatwal GS, Preissner KT, Müller-Berghaus G, Blobel H. Specific binding of the human S protein (vitronectin) to streptococci, Staphylococcus aureus, and Escherichia coli. Infect Immun. 1987 Aug;55(8):1878–1883. [Europe PMC free article] [Abstract] [Google Scholar]
  • de Boer HC, Preissner KT, Bouma BN, de Groot PG. Internalization of vitronectin-thrombin-antithrombin complex by endothelial cells leads to deposition of the complex into the subendothelial matrix. J Biol Chem. 1995 Dec 22;270(51):30733–30740. [Abstract] [Google Scholar]
  • Declerck PJ, De Mol M, Alessi MC, Baudner S, Pâques EP, Preissner KT, Müller-Berghaus G, Collen D. Purification and characterization of a plasminogen activator inhibitor 1 binding protein from human plasma. Identification as a multimeric form of S protein (vitronectin). J Biol Chem. 1988 Oct 25;263(30):15454–15461. [Abstract] [Google Scholar]
  • Elsinghorst EA. Measurement of invasion by gentamicin resistance. Methods Enzymol. 1994;236:405–420. [Abstract] [Google Scholar]
  • Fussenegger M, Kahrs AF, Facius D, Meyer TF. Tetrapac (tpc), a novel genotype of Neisseria gonorrhoeae affecting epithelial cell invasion, natural transformation competence and cell separation. Mol Microbiol. 1996 Mar;19(6):1357–1372. [Abstract] [Google Scholar]
  • Griffiss JM, Jarvis GA, O'Brien JP, Eads MM, Schneider H. Lysis of Neisseria gonorrhoeae initiated by binding of normal human IgM to a hexosamine-containing lipooligosaccharide epitope(s) is augmented by strain-specific, properdin-binding-dependent alternative complement pathway activation. J Immunol. 1991 Jul 1;147(1):298–305. [Abstract] [Google Scholar]
  • Handsfield HH. Disseminated gonococcal infection. Clin Obstet Gynecol. 1975 Mar;18(1):131–142. [Abstract] [Google Scholar]
  • Hayman EG, Pierschbacher MD, Suzuki S, Ruoslahti E. Vitronectin--a major cell attachment-promoting protein in fetal bovine serum. Exp Cell Res. 1985 Oct;160(2):245–258. [Abstract] [Google Scholar]
  • Hess S, Kanse SM, Kost C, Preissner KT. The versatility of adhesion receptor ligands in haemostasis: morpho-regulatory functions of vitronectin. Thromb Haemost. 1995 Jul;74(1):258–265. [Abstract] [Google Scholar]
  • Holmes KK, Counts GW, Beaty HN. Disseminated gonococcal infection. Ann Intern Med. 1971 Jun;74(6):979–993. [Abstract] [Google Scholar]
  • HUNTER WM, GREENWOOD FC. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. [Abstract] [Google Scholar]
  • Jarvis GA. Analysis of C3 deposition and degradation on Neisseria meningitidis and Neisseria gonorrhoeae. Infect Immun. 1994 May;62(5):1755–1760. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kahrs AF, Bihlmaier A, Facius D, Meyer TF. Generalized transposon shuttle mutagenesis in Neisseria gonorrhoeae: a method for isolating epithelial cell invasion-defective mutants. Mol Microbiol. 1994 Jun;12(5):819–831. [Abstract] [Google Scholar]
  • Kohl PK. Epidemiology of sexually transmitted diseases. What does it tell us? Sex Transm Dis. 1994 Mar-Apr;21(2 Suppl):S81–S83. [Abstract] [Google Scholar]
  • Kronvall G, Myhre EB, Björck L, Berggård I. Binding of aggregated human beta2-microglobulin to surface protein structure in group A, C, and G streptococci. Infect Immun. 1978 Oct;22(1):136–142. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kupsch EM, Knepper B, Kuroki T, Heuer I, Meyer TF. Variable opacity (Opa) outer membrane proteins account for the cell tropisms displayed by Neisseria gonorrhoeae for human leukocytes and epithelial cells. EMBO J. 1993 Feb;12(2):641–650. [Europe PMC free article] [Abstract] [Google Scholar]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [Abstract] [Google Scholar]
  • Laurière M. A semidry electroblotting system efficiently transfers both high- and low-molecular-weight proteins separated by SDS-PAGE. Anal Biochem. 1993 Jul;212(1):206–211. [Abstract] [Google Scholar]
  • Limper AH, Standing JE, Hoffman OA, Castro M, Neese LW. Vitronectin binds to Pneumocystis carinii and mediates organism attachment to cultured lung epithelial cells. Infect Immun. 1993 Oct;61(10):4302–4309. [Europe PMC free article] [Abstract] [Google Scholar]
  • Lydon MJ, Foulger CA. Cell-substratum interactions: serum spreading factor. Biomaterials. 1988 Nov;9(6):525–527. [Abstract] [Google Scholar]
  • Makino S, van Putten JP, Meyer TF. Phase variation of the opacity outer membrane protein controls invasion by Neisseria gonorrhoeae into human epithelial cells. EMBO J. 1991 Jun;10(6):1307–1315. [Europe PMC free article] [Abstract] [Google Scholar]
  • Milis L, Morris CA, Sheehan MC, Charlesworth JA, Pussell BA. Vitronectin-mediated inhibition of complement: evidence for different binding sites for C5b-7 and C9. Clin Exp Immunol. 1993 Apr;92(1):114–119. [Abstract] [Google Scholar]
  • Nowicki S, Martens MG, Nowicki BJ. Gonococcal infection in a nonhuman host is determined by human complement C1q. Infect Immun. 1995 Dec;63(12):4790–4794. [Europe PMC free article] [Abstract] [Google Scholar]
  • Panetti TS, McKeown-Longo PJ. Receptor-mediated endocytosis of vitronectin is regulated by its conformational state. J Biol Chem. 1993 Jun 5;268(16):11988–11993. [Abstract] [Google Scholar]
  • Panetti TS, McKeown-Longo PJ. The alpha v beta 5 integrin receptor regulates receptor-mediated endocytosis of vitronectin. J Biol Chem. 1993 Jun 5;268(16):11492–11495. [Abstract] [Google Scholar]
  • Panetti TS, Wilcox SA, Horzempa C, McKeown-Longo PJ. Alpha v beta 5 integrin receptor-mediated endocytosis of vitronectin is protein kinase C-dependent. J Biol Chem. 1995 Aug 4;270(31):18593–18597. [Abstract] [Google Scholar]
  • Petersen BH, Graham JA, Brooks GF. Human deficiency of the eighth component of complement. The requirement of C8 for serum Neisseria gonorrhoeae bactericidal activity. J Clin Invest. 1976 Feb;57(2):283–290. [Europe PMC free article] [Abstract] [Google Scholar]
  • Piot P, Islam MQ. Sexually transmitted diseases in the 1990s. Global epidemiology and challenges for control. Sex Transm Dis. 1994 Mar-Apr;21(2 Suppl):S7–13. [Abstract] [Google Scholar]
  • Porat N, Apicella MA, Blake MS. A lipooligosaccharide-binding site on HepG2 cells similar to the gonococcal opacity-associated surface protein Opa. Infect Immun. 1995 Jun;63(6):2164–2172. [Europe PMC free article] [Abstract] [Google Scholar]
  • Preissner KT, Grulich-Henn J, Ehrlich HJ, Declerck P, Justus C, Collen D, Pannekoek H, Müller-Berghaus G. Structural requirements for the extracellular interaction of plasminogen activator inhibitor 1 with endothelial cell matrix-associated vitronectin. J Biol Chem. 1990 Oct 25;265(30):18490–18498. [Abstract] [Google Scholar]
  • Richardson WP, Sadoff JC. Induced engulfment of Neisseria gonorrhoeae by tissue culture cells. Infect Immun. 1988 Sep;56(9):2512–2514. [Europe PMC free article] [Abstract] [Google Scholar]
  • Ringnér M, Valkonen KH, Wadström T. Binding of vitronectin and plasminogen to Helicobacter pylori. FEMS Immunol Med Microbiol. 1994 Jun;9(1):29–34. [Abstract] [Google Scholar]
  • Robertson BD, Frosch M, van Putten JP. The role of galE in the biosynthesis and function of gonococcal lipopolysaccharide. Mol Microbiol. 1993 May;8(5):891–901. [Abstract] [Google Scholar]
  • Schwan ET, Robertson BD, Brade H, van Putten JP. Gonococcal rfaF mutants express Rd2 chemotype LPS and do not enter epithelial host cells. Mol Microbiol. 1995 Jan;15(2):267–275. [Abstract] [Google Scholar]
  • Shaw JH, Falkow S. Model for invasion of human tissue culture cells by Neisseria gonorrhoeae. Infect Immun. 1988 Jun;56(6):1625–1632. [Europe PMC free article] [Abstract] [Google Scholar]
  • Simon D, Rest RF. Escherichia coli expressing a Neisseria gonorrhoeae opacity-associated outer membrane protein invade human cervical and endometrial epithelial cell lines. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5512–5516. [Europe PMC free article] [Abstract] [Google Scholar]
  • Stockmann A, Hess S, Declerck P, Timpl R, Preissner KT. Multimeric vitronectin. Identification and characterization of conformation-dependent self-association of the adhesive protein. J Biol Chem. 1993 Oct 25;268(30):22874–22882. [Abstract] [Google Scholar]
  • Sweet RL. Sexually transmitted diseases. Pelvic inflammatory disease and infertility in women. Infect Dis Clin North Am. 1987 Mar;1(1):199–215. [Abstract] [Google Scholar]
  • Sweet RL, Draper DL, Hadley WK. Etiology of acute salpingitis: influence of episode number and duration of symptoms. Obstet Gynecol. 1981 Jul;58(1):62–68. [Abstract] [Google Scholar]
  • Tomasini BR, Mosher DF. Vitronectin. Prog Hemost Thromb. 1991;10:269–305. [Abstract] [Google Scholar]
  • Tramont EC. Inhibition of adherence of Neisseria gonorrhoeae by human genital secretions. J Clin Invest. 1977 Jan;59(1):117–124. [Europe PMC free article] [Abstract] [Google Scholar]
  • Tran Van Nhieu G, Isberg RR. Isolation and identification of eukaryotic receptors promoting bacterial internalization. Methods Enzymol. 1994;236:307–318. [Abstract] [Google Scholar]
  • Ullberg M, Kuusela P, Kristiansen BE, Kronvall G. Binding of plasminogen to Neisseria meningitidis and Neisseria gonorrhoeae and formation of surface-associated plasmin. J Infect Dis. 1992 Dec;166(6):1329–1334. [Abstract] [Google Scholar]
  • Underwood PA, Bennett FA. A comparison of the biological activities of the cell-adhesive proteins vitronectin and fibronectin. J Cell Sci. 1989 Aug;93(Pt 4):641–649. [Abstract] [Google Scholar]
  • van Putten JP, Paul SM. Binding of syndecan-like cell surface proteoglycan receptors is required for Neisseria gonorrhoeae entry into human mucosal cells. EMBO J. 1995 May 15;14(10):2144–2154. [Europe PMC free article] [Abstract] [Google Scholar]
  • Virji M, Everson JS. Comparative virulence of opacity variants of Neisseria gonorrhoeae strain P9. Infect Immun. 1981 Mar;31(3):965–970. [Europe PMC free article] [Abstract] [Google Scholar]
  • Virji M, Makepeace K, Moxon ER. Distinct mechanisms of interactions of Opc-expressing meningococci at apical and basolateral surfaces of human endothelial cells; the role of integrins in apical interactions. Mol Microbiol. 1994 Oct;14(1):173–184. [Abstract] [Google Scholar]
  • Woods A, Couchman JR. Heparan sulfate proteoglycans and signalling in cell adhesion. Adv Exp Med Biol. 1992;313:87–96. [Abstract] [Google Scholar]
  • Woods A, Couchman JR. Protein kinase C involvement in focal adhesion formation. J Cell Sci. 1992 Feb;101(Pt 2):277–290. [Abstract] [Google Scholar]
Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

Full text links 

Read article at publisher's site: https://doi.org/10.1128/iai.65.9.3857-3866.1997

Read article for free, from open access legal sources, via Unpaywall: https://iai.asm.org/content/iai/65/9/3857.full.pdfUnpaywall

Free after 4 months at iai.asm.org
http://iai.asm.org/cgi/reprint/65/9/3857

Free to read at iai.asm.org
http://iai.asm.org/cgi/content/abstract/65/9/3857

Citations & impact 

Impact metrics

61
Citations
Jump to Citations

Citations of article over time

Alternative metrics

Altmetric item for https://www.altmetric.com/details/105785509
Altmetric
Discover the attention surrounding your research
https://www.altmetric.com/details/105785509

Smart citations by scite.ai
Smart citations by scite.ai include citation statements extracted from the full text of the citing article. The number of the statements may be higher than the number of citations provided by EuropePMC if one paper cites another multiple times or lower if scite has not yet processed some of the citing articles.
Explore citation contexts and check if this article has been supported or disputed.
https://scite.ai/reports/10.1128/iai.65.9.3857-3866.1997

Supporting
Mentioning
Contrasting
0
48
2

Article citations

Go to all (61) article citations

Similar Articles 

To arrive at the top five similar articles we use a word-weighted algorithm to compare words from the Title and Abstract of each citation.