title: Surface glycosaminoglycans mediate adherence between HeLa cells and Lactobacillus salivarius Lv72.
abstract: BACKGROUND
  The adhesion of lactobacilli to the vaginal surface is of paramount importance to develop their probiotic functions. For this reason, the role of HeLa cell surface proteoglycans in the attachment of Lactobacillus salivarius Lv72, a mutualistic strain of vaginal origin, was investigated.
  
  
  RESULTS
  Incubation of cultures with a variety of glycosaminoglycans (chondroitin sulfate A and C, heparin and heparan sulfate) resulted in marked binding interference. However, no single glycosaminoglycan was able to completely abolish cell binding, the sum of all having an additive effect that suggests cooperation between them and recognition of specific adhesins on the bacterial surface. In contrast, chondroitin sulfate B enhanced cell to cell attachment, showing the relevance of the stereochemistry of the uronic acid and the sulfation pattern on binding. Elimination of the HeLa surface glycosaminoglycans with lyases also resulted in severe adherence impairment. Advantage was taken of the Lactobacillus-glycosaminoglycans interaction to identify an adhesin from the bacterial surface. This protein, identify as a soluble binding protein of an ABC transporter system (OppA) by MALDI-TOF/(MS), was overproduced in Escherichia coli, purified and shown to interfere with L. salivarius Lv72 adhesion to HeLa cells.
  
  
  CONCLUSIONS
  These data suggest that glycosaminoglycans play a fundamental role in attachment of mutualistic bacteria to the epithelium that lines the cavities where the normal microbiota thrives, OppA being a bacterial adhesin involved in the process.
authors.affiliation: Área de Microbiología, Universidad de Oviedo, Julián Clavería 6 33006 Oviedo, Spain. quirosluis@uniovi.es.
authors.forename: Rebeca
authors.initials: R
authors.lastname: Martín
authors.forename: Carla
authors.initials: C
authors.lastname: Martín
authors.forename: Susana
authors.initials: S
authors.lastname: Escobedo
authors.forename: Juan E
authors.initials: JE
authors.lastname: Suárez
authors.forename: Luis M
authors.initials: LM
authors.lastname: Quirós
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism; D018829:Adhesins, Bacterial / Q000235:genetics / Q000378:metabolism*; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells / Q000737:chemistry* / Q000382:microbiology*; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D015870:Gene Expression; D006025:Glycosaminoglycans / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007778:Lactobacillus / Q000302:isolation & purification / Q000502:physiology*
chemical_list: D018528:ATP-Binding Cassette Transporters; D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; D006025:Glycosaminoglycans
references.citation: J Virol. 2001 Feb;75(3):1565-70
references.pmid: 11152531
references.citation: J Exp Med. 1995 Aug 1;182(2):511-7
references.pmid: 7629509
references.citation: J Biol Chem. 1998 Sep 25;273(39):24979-82
references.pmid: 9737951
references.citation: J Cell Sci. 1990 Nov;97 ( Pt 3):411-7
references.pmid: 1705939
references.citation: Curr Opin Cell Biol. 1990 Oct;2(5):815-20
references.pmid: 2083082
references.citation: J Clin Invest. 2001 Jul;108(2):169-73
references.pmid: 11457867
references.citation: Nature. 2001 Dec 6;414(6864):648-52
references.pmid: 11740562
references.citation: Infect Immun. 2000 Apr;68(4):1884-92
references.pmid: 10722578
references.citation: J Clin Invest. 2000 Oct;106(8):995-1002
references.pmid: 11032859
references.citation: Infect Immun. 2002 Mar;70(3):1530-7
references.pmid: 11854241
references.citation: J Cell Sci. 2000 Jan;113 Pt 2:193-205
references.pmid: 10633071
references.citation: BMC Microbiol. 2011 Aug 19;11:185
references.pmid: 21854595
references.citation: Annu Rev Biochem. 1999;68:729-77
references.pmid: 10872465
references.citation: J Cell Biol. 1987 Jun;104(6):1683-91
references.pmid: 3584245
references.citation: J Med Microbiol. 1992 Jul;37(1):38-42
references.pmid: 1625314
references.citation: Nature. 1970 Aug 15;227(5259):680-5
references.pmid: 5432063
references.citation: Infect Immun. 1995 Sep;63(9):3520-6
references.pmid: 7642286
references.citation: Chemphyschem. 2009 Jul 13;10(9-10):1672-5
references.pmid: 19475637
references.citation: Cell Mol Life Sci. 2009 Nov;66(21):3421-34
references.pmid: 19629389
references.citation: Cell. 1991 Jun 28;65(7):1099-102
references.pmid: 1905978
references.citation: J AOAC Int. 2012 Jan-Feb;95(1):31-4
references.pmid: 22468339
references.citation: J Biotechnol. 2007 Apr 30;129(2):290-307
references.pmid: 17337080
references.citation: J Infect Dis. 1995 May;171(5):1237-43
references.pmid: 7751699
references.citation: Glycobiology. 2000 Nov;10(11):1147-56
references.pmid: 11087707
references.citation: Microbes Infect. 2006 Feb;8(2):316-22
references.pmid: 16239116
references.citation: Proteomics. 2008 Apr;8(7):1415-28
references.pmid: 18306176
references.citation: FEMS Microbiol Lett. 2012 Mar;328(2):166-73
references.pmid: 22224921
references.citation: Cell Tissue Res. 2010 Jan;339(1):237-46
references.pmid: 19513755
references.citation: Infect Immun. 2006 Jan;74(1):435-41
references.pmid: 16368999
references.citation: J Appl Microbiol. 2007 Feb;102(2):442-51
references.pmid: 17241350
references.citation: Shock. 2005 Dec;24(6):571-6
references.pmid: 16317389
references.citation: Mol Microbiol. 1993 Aug;9(4):687-94
references.pmid: 7901732
references.citation: J Bacteriol. 1999 Aug;181(16):4873-8
references.pmid: 10438757
references.citation: FEMS Microbiol Lett. 1999 Aug 15;177(2):313-7
references.pmid: 10474198
references.citation: J Appl Microbiol. 2010 Mar;108(3):1050-1059
references.pmid: 19735320
references.citation: Infect Immun. 2000 Jul;68(7):4005-11
references.pmid: 10858215
references.citation: Mol Microbiol. 1999 Apr;32(2):233-42
references.pmid: 10231481
references.citation: Infect Immun. 1998 May;66(5):1985-9
references.pmid: 9573080
references.citation: EMBO J. 2009 May 6;28(9):1332-40
references.pmid: 19300437
references.citation: Proc Natl Acad Sci U S A. 2011 Mar 15;108 Suppl 1:4680-7
references.pmid: 20534435
references.citation: Virology. 2001 Apr 10;282(2):245-55
references.pmid: 11289807
references.citation: Int Microbiol. 2008 Dec;11(4):261-6
references.pmid: 19204898
references.citation: Microb Pathog. 2006 Mar;40(3):126-32
references.pmid: 16455229
references.citation: Biochimie. 2012 Jun;94(6):1291-9
references.pmid: 22386869
references.citation: Immunity. 2003 Jan;18(1):27-39
references.pmid: 12530973
references.citation: J Infect Dis. 2001 Jul 15;184(2):181-7
references.pmid: 11424015
references.citation: PLoS One. 2010 Aug 17;5(8):e12225
references.pmid: 20808924
references.citation: Arch Microbiol. 2002 Jun;177(6):457-67
references.pmid: 12029391
references.citation: J Cell Biol. 1993 Nov;123(3):759-66
references.pmid: 8227137
references.citation: PLoS One. 2009;4(1):e4166
references.pmid: 19132104
references.citation: Infect Immun. 1997 Jan;65(1):78-88
references.pmid: 8975895
references.citation: FEMS Immunol Med Microbiol. 2008 Oct;54(1):1-17
references.pmid: 18631181
references.citation: Enferm Infecc Microbiol Clin. 2008 Mar;26(3):160-7
references.pmid: 18358215
references.citation: J Cell Biol. 1986 Dec;103(6 Pt 2):2683-96
references.pmid: 3025223
references.citation: Cell Host Microbe. 2007 Nov 15;2(5):316-27
references.pmid: 18005753
references.citation: Med Microbiol Immunol. 2006 Sep;195(3):133-41
references.pmid: 16378213
references.citation: J Med Microbiol. 2001 Feb;50(2):183-190
references.pmid: 11211227
references.citation: G3 (Bethesda). 2013 Aug 07;3(8):1213-24
references.pmid: 23550136
references.citation: Trends Cell Biol. 2001 Feb;11(2):75-82
references.pmid: 11166215
references.citation: Biosci Biotechnol Biochem. 2006 Jul;70(7):1622-8
references.pmid: 16861796
references.citation: J Appl Microbiol. 1997 Oct;83(4):413-20
references.pmid: 9351223
references.citation: J Infect Dis. 1996 Nov;174(5):1058-63
references.pmid: 8896509
references.citation: FEMS Microbiol Lett. 2007 Nov;276(2):140-8
references.pmid: 17888009
references.citation: Cell Microbiol. 2004 May;6(5):435-45
references.pmid: 15056214
references.citation: Probiotics Antimicrob Proteins. 2011 Mar;3(1):15-20
references.pmid: 26781495
references.citation: Mol Microbiol. 1998 Mar;27(6):1203-12
references.pmid: 9570405
references.citation: Infect Immun. 2009 Dec;77(12):5528-36
references.pmid: 19805539
references.citation: J Biol Chem. 2009 Jul 3;284(27):18194-201
references.pmid: 19395379