#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Bacteriophage Lysin Mediates the Binding of to Human Platelets through Interaction with Fibrinogen


The binding of bacteria to human platelets is a likely central mechanism in the pathogenesis of infective endocarditis. We have previously found that platelet binding by Streptococcus mitis SF100 is mediated by surface components encoded by a lysogenic bacteriophage, SM1. We now demonstrate that SM1-encoded lysin contributes to platelet binding via its direct interaction with fibrinogen. Far Western blotting of platelets revealed that fibrinogen was the major membrane-associated protein bound by lysin. Analysis of lysin binding with purified fibrinogen in vitro confirmed that these proteins could bind directly, and that this interaction was both saturable and inhibitable. Lysin bound both the Aα and Bβ chains of fibrinogen, but not the γ subunit. Binding of lysin to the Bβ chain was further localized to a region within the fibrinogen D fragment. Disruption of the SF100 lysin gene resulted in an 83±3.1% reduction (mean ± SD) in binding to immobilized fibrinogen by this mutant strain (PS1006). Preincubation of this isogenic mutant with purified lysin restored fibrinogen binding to wild type levels. When tested in a co-infection model of endocarditis, loss of lysin expression resulted in a significant reduction in virulence, as measured by achievable bacterial densities (CFU/g) within vegetations, kidneys, and spleens. These results indicate that bacteriophage-encoded lysin is a multifunctional protein, representing a new class of fibrinogen-binding proteins. Lysin appears to be cell wall-associated through its interaction with choline. Once on the bacterial surface, lysin can bind fibrinogen directly, which appears to be an important interaction for the pathogenesis of endocarditis.


Vyšlo v časopise: Bacteriophage Lysin Mediates the Binding of to Human Platelets through Interaction with Fibrinogen. PLoS Pathog 6(8): e32767. doi:10.1371/journal.ppat.1001047
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1001047

Souhrn

The binding of bacteria to human platelets is a likely central mechanism in the pathogenesis of infective endocarditis. We have previously found that platelet binding by Streptococcus mitis SF100 is mediated by surface components encoded by a lysogenic bacteriophage, SM1. We now demonstrate that SM1-encoded lysin contributes to platelet binding via its direct interaction with fibrinogen. Far Western blotting of platelets revealed that fibrinogen was the major membrane-associated protein bound by lysin. Analysis of lysin binding with purified fibrinogen in vitro confirmed that these proteins could bind directly, and that this interaction was both saturable and inhibitable. Lysin bound both the Aα and Bβ chains of fibrinogen, but not the γ subunit. Binding of lysin to the Bβ chain was further localized to a region within the fibrinogen D fragment. Disruption of the SF100 lysin gene resulted in an 83±3.1% reduction (mean ± SD) in binding to immobilized fibrinogen by this mutant strain (PS1006). Preincubation of this isogenic mutant with purified lysin restored fibrinogen binding to wild type levels. When tested in a co-infection model of endocarditis, loss of lysin expression resulted in a significant reduction in virulence, as measured by achievable bacterial densities (CFU/g) within vegetations, kidneys, and spleens. These results indicate that bacteriophage-encoded lysin is a multifunctional protein, representing a new class of fibrinogen-binding proteins. Lysin appears to be cell wall-associated through its interaction with choline. Once on the bacterial surface, lysin can bind fibrinogen directly, which appears to be an important interaction for the pathogenesis of endocarditis.


Zdroje

1. KnoxKW

HunterN

1991 The role of oral bacteria in the pathogenesis of infective endocarditis. Aust Dent J 36 286 292

2. BashoreTM

CabellC

FowlerVJr

2006 Update on infective endocarditis. Curr Probl Cardiol 31 274 352

3. GaneshVK

RiveraJJ

SmedsE

KoYP

BowdenMG

2008 A structural model of the Staphylococcus aureus ClfA-fibrinogen interaction opens new avenues for the design of anti-staphylococcal therapeutics. PLoS Pathog 4 e1000226

4. McDevittD

NanavatyT

House-PompeoK

BellE

TurnerN

1997 Characterization of the interaction between the Staphylococcus aureus clumping factor (ClfA) and fibrinogen. Eur J Biochem 247 416 424

5. MiajlovicH

LoughmanA

BrennanM

CoxD

FosterTJ

2007 Both complement- and fibrinogen-dependent mechanisms contribute to platelet aggregation mediated by Staphylococcus aureus clumping factor B. Infect Immun 75 3335 3343

6. FitzgeraldJR

FosterTJ

CoxD

2006 The interaction of bacterial pathogens with platelets. Nat Rev Microbiol 4 445 457

7. LoughmanA

FitzgeraldJR

BrennanMP

HigginsJ

DownerR

2005 Roles for fibrinogen, immunoglobulin and complement in platelet activation promoted by Staphylococcus aureus clumping factor A. Mol Microbiol 57 804 818

8. O'BrienL

KerriganSW

KawG

HoganM

PenadesJ

2002 Multiple mechanisms for the activation of human platelet aggregation by Staphylococcus aureus: roles for the clumping factors ClfA and ClfB, the serine-aspartate repeat protein SdrE and protein A. Mol Microbiol 44 1033 1044

9. FordI

DouglasCW

1997 The role of platelets in infective endocarditis. Platelets 8 285 294

10. Bek-ThomsenM

TettelinH

HanceI

NelsonKE

KilianM

2008 Population diversity and dynamics of Streptococcus mitis, Streptococcus oralis, and Streptococcus infantis in the upper respiratory tracts of adults, determined by a nonculture strategy. Infect Immun 76 1889 1896

11. NgKH

LeeS

YipSF

QueTL

2005 A case of Streptococcus mitis endocarditis successfully treated by linezolid. Hong Kong Med J 11 411 413

12. HuangIF

ChiouCC

LiuYC

HsiehKS

2002 Endocarditis caused by penicillin-resistant Streptococcus mitis in a 12-year-old boy. J Microbiol Immunol Infect 35 129 132

13. HallGE

BaddourLM

2002 Apparent failure of endocarditis prophylaxis caused by penicillin-resistant Streptococcus mitis. Am J Med Sci 324 51 53

14. ChayakulP

HortiwakulR

YipintsoiT

IngviyaN

2002 Varidans streptococci in the oral flora of the patients at risk for infective endocarditis: species and penicillin susceptibilities. J Med Assoc Thai 85 825 830

15. MitchellJ

SullamPM

2009 Streptococcus mitis phage-encoded adhesins mediate attachment to {alpha}2-8-linked sialic acid residues on platelet membrane gangliosides. Infect Immun 77 3485 3490

16. MitchellJ

SibooIR

TakamatsuD

ChambersHF

SullamPM

2007 Mechanism of cell surface expression of the Streptococcus mitis platelet binding proteins PblA and PblB. Mol Microbiol 64 844 857

17. BensingBA

SibooIR

SullamPM

2001 Proteins PblA and PblB of Streptococcus mitis, which promote binding to human platelets, are encoded within a lysogenic bacteriophage. Infect Immun 69 6186 6192

18. Marchler-BauerA

PanchenkoAR

ShoemakerBA

ThiessenPA

GeerLY

2002 CDD: a database of conserved domain alignments with links to domain three-dimensional structure. Nucleic Acids Res 30 281 283

19. SheehanMM

GarciaJL

LopezR

GarciaP

1997 The lytic enzyme of the pneumococcal phage Dp-1: a chimeric lysin of intergeneric origin. Mol Microbiol 25 717 725

20. PritchardDG

DongS

KirkMC

CarteeRT

BakerJR

2007 LambdaSa1 and LambdaSa2 prophage lysins of Streptococcus agalactiae. Appl Environ Microbiol 73 7150 7154

21. CaubinJ

MartinH

RoaA

CosanoI

PozueloM

2001 Choline-binding domain as a novel affinity tag for purification of fusion proteins produced in Pichia pastoris. Biotechnol Bioeng 74 164 171

22. Ni EidhinD

PerkinsS

FrancoisP

VaudauxP

HookM

1998 Clumping factor B (ClfB), a new surface-located fibrinogen-binding adhesin of Staphylococcus aureus. Mol Microbiol 30 245 257

23. WalshEJ

MiajlovicH

GorkunOV

FosterTJ

2008 Identification of the Staphylococcus aureus MSCRAMM clumping factor B (ClfB) binding site in the alphaC-domain of human fibrinogen. Microbiology 154 550 558

24. GeorgeNP

WeiQ

ShinPK

KonstantopoulosK

RossJM

2006 Staphylococcus aureus adhesion via Spa, ClfA, and SdrCDE to immobilized platelets demonstrates shear-dependent behavior. Arterioscler Thromb Vasc Biol 26 2394 2400

25. FitzgeraldJR

LoughmanA

KeaneF

BrennanM

KnobelM

2006 Fibronectin-binding proteins of Staphylococcus aureus mediate activation of human platelets via fibrinogen and fibronectin bridges to integrin GPIIb/IIIa and IgG binding to the FcgammaRIIa receptor. Mol Microbiol 59 212 230

26. SimpsonKH

BowdenG

HookM

AnvariB

2003 Measurement of adhesive forces between individual Staphylococcus aureus MSCRAMMs and protein-coated surfaces by use of optical tweezers. J Bacteriol 185 2031 2035

27. LeeSY

KimKK

ChoeSJ

2001 Binding of oral streptococci to human fibrinogen. Oral Microbiol Immunol 16 88 93

28. ReuterswardA

MiornerH

WagnerM

KronvallG

1985 Variations in binding of mammalian fibrinogens to streptococci groups A, B, C, E, G and to Staphylococcus aureus. Acta Pathol Microbiol Immunol Scand B 93 77 82

29. KuuselaP

VartioT

VuentoM

MyhreEB

1985 Attachment of staphylococci and streptococci on fibronectin, fibronectin fragments, and fibrinogen bound to a solid phase. Infect Immun 50 77 81

30. PietrocolaG

SchubertA

VisaiL

TortiM

FitzgeraldJR

2005 FbsA, a fibrinogen-binding protein from Streptococcus agalactiae, mediates platelet aggregation. Blood 105 1052 1059

31. HerzbergMC

NobbsA

TaoL

KilicA

BeckmanE

2005 Oral streptococci and cardiovascular disease: searching for the platelet aggregation-associated protein gene and mechanisms of Streptococcus sanguis-induced thrombosis. J Periodontol 76 2101 2105

32. KerriganSW

DouglasI

WrayA

HeathJ

ByrneMF

2002 A role for glycoprotein Ib in Streptococcus sanguis-induced platelet aggregation. Blood 100 509 516

33. DouglasCW

BrownPR

PrestonFE

1990 Platelet aggregation by oral streptococci. FEMS Microbiol Lett 60 63 67

34. CourtneyHS

DaleJB

HastyDL

2002 Mapping the fibrinogen-binding domain of serum opacity factor of group A streptococci. Curr Microbiol 44 236 240

35. HeilmannC

HartleibJ

HussainMS

PetersG

2005 The multifunctional Staphylococcus aureus autolysin Aaa mediates adherence to immobilized fibrinogen and fibronectin. Infect Immun 73 4793 4802

36. HeilmannC

ThummG

ChhatwalGS

HartleibJ

UekotterA

2003 Identification and characterization of a novel autolysin (Aae) with adhesive properties from Staphylococcus epidermidis. Microbiology 149 2769 2778

37. BrouilletteE

LacasseP

ShkretaL

BelangerJ

GrondinG

2002 DNA immunization against the clumping factor A (ClfA) of Staphylococcus aureus. Vaccine 20 2348 2357

38. EntenzaJM

MoreillonP

SennMM

KormanecJ

DunmanPM

2005 Role of sigmaB in the expression of Staphylococcus aureus cell wall adhesins ClfA and FnbA and contribution to infectivity in a rat model of experimental endocarditis. Infect Immun 73 990 998

39. SullamPM

ValoneFH

MillsJ

1987 Mechanisms of platelet aggregation by viridans group streptococci. Infect Immun 55 1743 1750

40. DhawanVK

YeamanMR

CheungAL

KimE

SullamPM

1997 Phenotypic resistance to thrombin-induced platelet microbicidal protein in vitro is correlated with enhanced virulence in experimental endocarditis due to Staphylococcus aureus. Infect Immun 65 3293 3299

41. CheungAL

EberhardtKJ

ChungE

YeamanMR

SullamPM

1994 Diminished virulence of a sar-/agr- mutant of Staphylococcus aureus in the rabbit model of endocarditis. J Clin Invest 94 1815 1822

42. AC

SGR

IL

JO

2003 Bacteria-binding plasma proteins in pellicles formed on hydroxyapatite in vitro and on teeth in vivo. Oral Microbiology Immunology 18 203 207

43. NagataH

MurakamiY

InoshitaE

ShizukuishiS

TsunemitsuA

1990 Inhibitory effect of human plasma andsaliva on co-aggregation between Bacteroides gingivalis and Streptococcus mitis. J Dent REs 69 1476 1479

44. YasminA

KennyJG

ShankarJ

DarbyAC

HallN

2010 Comparative genomics and transduction potential of Enterococcus faecalis temperate bacteriophages. J Bacteriol 192 1122 1130

45. RomeroP

CroucherNJ

HillerNL

HuFZ

EhrlichGD

2009 Comparative genomic analysis of ten Streptococcus pneumoniae temperate bacteriophages. J Bacteriol 191 4854 4862

46. ChaffinDO

C.E.R

1998 Blue/white screening of recombinant plasmids in Gram-positive bacteria by interruption of alkaline phosphatase gene (phoZ) expression. Gene 219 91 99

47. van de RijnI

KesslerRE

1980 Growth characteristics of group A streptococci in a new chemically defined medium. Infect Immun 27 444 448

48. DillardJP

VanderseaMW

YotherJ

1995 Characterization of the cassette containing genes for type 3 capsular polysaccharide biosynthesis in Streptococcus pneumoniae. J Exp Med 181 973 983

49. YotherJ

LeopoldK

WhiteJ

FischerW

1998 Generation and properties of a Streptococcus pneumoniae mutant which does not require choline or analogs for growth. J Bacteriol 180 2093 2101

50. HarmonJT

GrecoNJ

JamiesonGA

1992 Isolation of human platelet plasma membranes by glycerol lysis. Methods Enzymol 215 32 36

51. SibooIR

ChambersHF

SullamPM

2005 Role of SraP, a Serine-Rich Surface Protein of Staphylococcus aureus, in binding to human platelets. Infect Immun 73 2273 2280

52. SeoHS

CarteeRT

PritchardDG

NahmMH

2008 A new model of pneumococcal lipoteichoic acid structure resolves biochemical, biosynthetic, and serologic inconsistencies of the current model. J Bacteriol 190 2379 2387

53. SeoHS

MichalekSM

NahmMH

2008 Lipoteichoic acid is important in innate immune responses to Gram-positive bacteria. Infect Immun 76 206 213

54. XiongYQ

BensingBA

BayerAS

ChambersHF

SullamPM

2008 Role of the serine-rich surface glycoprotein GspB of Streptococcus gordonii in the pathogenesis of infective endocarditis. Microb Pathog 45 297 301

55. GianfaldoniC

CensiniS

HilleringmannM

MoschioniM

FacciottiC

2007 Streptococcus pneumoniae pilus subunits protect mice against lethal challenge. Infect Immun 75 1059 1062

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2010 Číslo 8
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Aktuální možnosti diagnostiky a léčby litiáz
nový kurz
Autori: MUDr. Tomáš Ürge, PhD.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#