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Thromb Haemost 2012; 108(02): 236-246
DOI: 10.1160/TH12-01-0032
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Streptococcus gordonii FSS2 Challisin affects fibrin clot formation by digestion of the αC region and cleavage of the N-terminal region of the Bβ chains of fibrinogen

Derek W. S. Harty
1   Institute of Dental Research, Westmead Millennium Institute and Westmead Centre for Oral Health, Sydney, Australia
,
Ramin M. Farahani
1   Institute of Dental Research, Westmead Millennium Institute and Westmead Centre for Oral Health, Sydney, Australia
2   Faculty of Dentistry, University of Sydney, Sydney, Australia
,
Mary R. Simonian
1   Institute of Dental Research, Westmead Millennium Institute and Westmead Centre for Oral Health, Sydney, Australia
,
Luke Hunter
3   School of Chemistry, University of New South Wales, Sydney, Australia
,
Neil Hunter
1   Institute of Dental Research, Westmead Millennium Institute and Westmead Centre for Oral Health, Sydney, Australia
2   Faculty of Dentistry, University of Sydney, Sydney, Australia
› Author Affiliations Financial support: This work was supported by Westmead Centre for Oral Health.
Further Information

Publication History

Received: 20 January 2012

Accepted after major revision: 17 April 2012

Publication Date:
25 November 2017 (online)

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Summary

Bacteria within endocarditis vegetations are encased in fibrin matrix that is resistant to resolution. We have previously shown that FSS2 Challisin, a serine protease from Streptococcus gordonii, is able to hydrolyse the Aα and Bβ chains of fibrinogen and has potent angiotensin converting enzyme (ACE) activity. The alteration in the structure of fibrin formed from FSS2 Challisin-degraded fibrinogen may therefore contribute to the resistant fibrin matrix. To this end, we have investigated the specific interactions of FSS2 Challisin with fibrinogen. FSS2 Challisin extensively degrades the αC region of fibrinogen Aα chains, hydrolysing both the αC-domain and αC-connnector. Additionally, the N-terminal region of the Bβ chains is cleaved twice, at Leu19 and Ser28, removing the B fibrinopeptides and ‘B’ knobs. Substrate analysis indicates FSS2 Challisin has specific requirement for proline two residues before the cleavage point and a neutral or basic un-branched amino acid preceding the cleavage point. Fibrin formation by thrombin was modified and the initiation of fibrinolysis extended, in FSS2 Challisin-treated plasma clots. Digestion of fibrinogen by FSS2 Challisin prior to thrombin action increased fiber density and fiber branch point density. The velocity of fibrinolysis was significantly slower for fibrin formed from FSS2 Challisin-treated fibrinogen but was faster when data was normalised for the increased fibrin density. Thromboelastography of whole blood treated with FSS2 Challisin indicated reduced clot coagulation time and increased shear resistance. Combined ACE and fibrinogenase activities of FSS2 Challisin suggest a pro-coagulant effect of this virulence factor which is conserved in the viridans streptococci.

Keywords

Fibrinogen - fibrin - thrombus - Streptococcus gordonii - infective endocarditis

 

  • References

  • 1 Mosesson MW. Fibrinogen and fibrin structure and functions. J Thromb Haemost 2005; 03: 1894-1904.
    CrossrefPubMedGoogle Scholar
  • 2 Doolittle RF, Kollman JM. Natively unfolded regions of the vertebrate fibrinogen molecule. Proteins 2006; 63: 391-397.
    CrossrefPubMedGoogle Scholar
  • 3 Weisel JW. Fibrinogen and fibrin. Adv Protein Chem 2005; 70: 247-299.
    PubMedGoogle Scholar
  • 4 Everse SJ, Spraggon G, Veerapandian L. et al. Crystal structure of fragment double-D from human fibrin with two different bound ligands. Biochemistry 1998; 37: 8637-8642.
    CrossrefPubMedGoogle Scholar
  • 5 Spraggon G, Everse SJ, Doolittle RF. Crystal structures of fragment D from human fibrinogen and its crosslinked counterpart from fibrin. Nature 1997; 389: 455-462.
    CrossrefPubMedGoogle Scholar
  • 6 Doolittle RF. Determining the crystal structure of fibrinogen. J Thromb Haemost 2004; 02: 683-689.
    CrossrefPubMedGoogle Scholar
  • 7 Kollman JM, Pandi L, Sawaya MR. et al. Crystal structure of human fibrinogen. Biochemistry 2009; 48: 3877-3886.
    CrossrefPubMedGoogle Scholar
  • 8 Laudano AP, Doolittle RF. Synthetic peptide derivatives that bind to fibrinogen and prevent the polymerization of fibrin monomers. Proc Natl Acad Sci USA 1978; 75: 3085-3089.
    CrossrefPubMedGoogle Scholar
  • 9 Weisel JW. Which knobs fit into which holes in fibrin polymerization?. J Thromb Haemost 2007; 05: 2340-2343.
    CrossrefPubMedGoogle Scholar
  • 10 Weisel JW, Veklich Y, Gorkun O. The sequence of cleavage of fibrinopeptides from fibrinogen is important for protofibril formation and enhancement of lateral aggregation in fibrin clots. J Mol Biol 1993; 232: 285-297.
    CrossrefPubMedGoogle Scholar
  • 11 Okumura N, Gorkun OV, Lord ST. Severely impaired polymerization of recombinant fibrinogen gamma-364 Asp --> His, the substitution discovered in a heterozygous individual. J Biol Chem 1997; 272: 29596-29601.
    CrossrefPubMedGoogle Scholar
  • 12 Litvinov RI, Gorkun OV, Owen SF. et al. Polymerization of fibrin: specificity, strength, and stability of knob-hole interactions studied at the single-molecule level. Blood 2005; 106: 2944-2951.
    CrossrefPubMedGoogle Scholar
  • 13 Litvinov RI, Gorkun OV, Galanakis DK. et al. Polymerization of fibrin: Direct observation and quantification of individual B: b knob-hole interactions. Blood 2007; 109: 130-138.
    CrossrefPubMedGoogle Scholar
  • 14 Chernysh IN, Nagaswami C, Weisel JW. Visualization and identification of the structures formed during early stages of fibrin polymerization. Blood 2011; 117: 4609-4614.
    CrossrefPubMedGoogle Scholar
  • 15 Galanakis DK. To gel or not to gel. Blood 2011; 117: 4406-4407.
    CrossrefPubMedGoogle Scholar
  • 16 Koh DC, Armugam A, Jeyaseelan K. Snake venom components and their applications in biomedicine. Cell Mol Life Sci 2006; 63: 3030-3041.
    CrossrefPubMedGoogle Scholar
  • 17 Weisel JW. Fibrin assembly. Lateral aggregation and the role of the two pairs of fi-brinopeptides. Biophys J 1986; 50: 1079-1093.
    CrossrefPubMedGoogle Scholar
  • 18 Doolittle RF, Pandi L. Binding of synthetic B knobs to fibrinogen changes the character of fibrin and inhibits its ability to activate tissue plasminogen activator and its destruction by plasmin. Biochemistry 2006; 45: 2657-2667.
    CrossrefPubMedGoogle Scholar
  • 19 Collet JP, Moen JL, Veklich YI. et al. The alphaC domains of fibrinogen affect the structure of the fibrin clot, its physical properties, and its susceptibility to fibrinolysis. Blood 2005; 106: 3824-3830.
    CrossrefPubMedGoogle Scholar
  • 20 Tsurupa G, Medved L. Fibrinogen alpha C domains contain cryptic plasminogen and tPA binding sites. Ann NY Acad Sci 2001; 936: 328-330.
    PubMedGoogle Scholar
  • 21 Tsurupa G, Medved L. Identification and characterization of novel tPA- and plasminogen-binding sites within fibrin(ogen) alpha C-domains. Biochemistry 2001; Jan 23 40: 801-808.
    CrossrefPubMedGoogle Scholar
  • 22 Gorkun OV, Veklich YI, Medved LV. et al. Role of the alpha C domains of fibrin in clot formation. Biochemistry 1994; Jun 7 33: 6986-6997.
    CrossrefPubMedGoogle Scholar
  • 23 Gorkun OV, Henschen-Edman AH, Ping LF. et al. Analysis of A alpha 251 fibrinogen: the alpha C domain has a role in polymerization, albeit more subtle than anticipated from the analogous proteolytic fragment X. Biochemistry 1998; 37: 15434-15441.
    CrossrefPubMedGoogle Scholar
  • 24 Medved L, Tsurupa G, Yakovlev S. Conformational changes upon conversion of fibrinogen into fibrin. The mechanisms of exposure of cryptic sites. Ann NY Acad Sci 2001; 936: 185-204.
    PubMedGoogle Scholar
  • 25 Litvinov RI, Yakovlev S, Tsurupa G. et al. Direct evidence for specific interactions of the fibrinogen alphaC-domains with the central E region and with each other. Biochemistry 2007; 46: 9133-9142.
    CrossrefPubMedGoogle Scholar
  • 26 Ping L, Huang L, Cardinali B. et al. Substitution of the human alphaC region with the analogous chicken domain generates a fibrinogen with severely impaired lateral aggregation: fibrin monomers assemble into protofibrils but protofibrils do not assemble into fibers. Biochemistry 2011; 50: 9066-9075.
    CrossrefPubMedGoogle Scholar
  • 27 Tsurupa G, Mahid A, Veklich Y. et al. Structure, stability, and interaction of fibrin alphaC-domain polymers. Biochemistry 2011; 50: 8028-8037.
    CrossrefPubMedGoogle Scholar
  • 28 Weisel JW, Medved L. The structure and function of the alpha C domains of fibrinogen. Ann NY Acad Sci 2001; 936: 312-327.
    PubMedGoogle Scholar
  • 29 Que YA, Moreillon P. Infective endocarditis. Nat Rev Cardiol 2011; 08: 322-336.
    CrossrefPubMedGoogle Scholar
  • 30 Harty DW, Hunter N. Carboxypeptidase activity common to viridans group streptococci cleaves angiotensin I to angiotensin II: an activity homologous to angiotensin-converting enzyme (ACE). Microbiology 2011; 157: 2143-2151.
    CrossrefPubMedGoogle Scholar
  • 31 Bonifait L, Vaillancourt K, Gottschalk M. et al. Purification and characterization of the subtilisin-like protease of Streptococcus suis that contributes to its virulence. Vet Microbiol 2011; 148: 333-340.
    CrossrefPubMedGoogle Scholar
  • 32 Harty DW, Patrikakis M, Hume EB. et al. The aggregation of human platelets by Lactobacillus species. J Gen Microbiol 1993; 139: 2945-2951.
    CrossrefPubMedGoogle Scholar
  • 33 Herzberg MC. Platelet-streptococcal interactions in endocarditis. Crit Rev Oral Biol Med 1996; 07: 222-236.
    CrossrefPubMedGoogle Scholar
  • 34 Herzberg MC. Coagulation and thrombosis in cardiovascular disease: plausible contributions of infectious agents. Ann Periodontol 2001; 06: 16-19.
    CrossrefPubMedGoogle Scholar
  • 35 Petersen HJ, Keane C, Jenkinson HF. et al. Human platelets recognize a novel surface protein, PadA, on Streptococcus gordonii through a unique interaction involving fibrinogen receptor GPIIbIIIa. Infect Immun 2010; 78: 413-422.
    CrossrefPubMedGoogle Scholar
  • 36 Nelson D, Goldstein JM, Boatright K. et al. pH-regulated secretion of a glyceraldehyde-3-phosphate dehydrogenase from Streptococcus gordonii FSS2: purification, characterization, and cloning of the gene encoding this enzyme. J Dent Res 2001; 80: 371-377.
    CrossrefPubMedGoogle Scholar
  • 37 Harty DW, Mayo JA, Cook SL. et al. Environmental regulation of glycosidase and peptidase production by Streptococcus gordonii FSS2. Microbiology 2000; 146: 1923-1931.
    CrossrefPubMedGoogle Scholar
  • 38 Gersh KC, Nagaswami C, Weisel JW. Fibrin network structure and clot mechanical properties are altered by incorporation of erythrocytes. Thromb Haemost 2009; 102: 1169-1175.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 39 Varju I, Sotonyi P, Machovich R. et al. Hindered dissolution of fibrin formed under mechanical stress. J Thromb Haemost 2011; 09: 979-986.
    CrossrefPubMedGoogle Scholar
  • 40 Carey RM, Siragy HM. Newly recognized components of the renin-angiotensin system: potential roles in cardiovascular and renal regulation. Endocr Rev 2003; 24: 261-271.
    CrossrefPubMedGoogle Scholar
  • 41 Jagroop IA, Mikhailidis DP. Angiotensin II can induce and potentiate shape change in human platelets: effect of losartan. J Hum Hypertens 2000; 14: 581-585.
    CrossrefPubMedGoogle Scholar
  • 42 Senchenkova EY, Russell J, Almeida-Paula LD. et al. Angiotensin II-mediated microvascular thrombosis. Hypertension 2010; 56: 1089-1095.
    CrossrefPubMedGoogle Scholar
  • 43 Watanabe T, Barker TA, Berk BC. Angiotensin II and the endothelium: diverse signals and effects. Hypertension 2005; 45: 163-169.
    CrossrefPubMedGoogle Scholar
  • 44 Manning JE, Hume EB, Hunter N. et al. An appraisal of the virulence factors associated with streptococcal endocarditis. J Med Microbiol 1994; 40: 110-114.
    CrossrefPubMedGoogle Scholar
  • 45 Cox D, Kerrigan SW, Watson SP. Platelets and the innate immune system: mechanisms of bacterial-induced platelet activation. J Thromb Haemost 2011; 09: 1097-1107.
    CrossrefPubMedGoogle Scholar
  • 46 Bensing BA, Lopez JA, Sullam PM. The Streptococcus gordonii surface proteins GspB and Hsa mediate binding to sialylated carbohydrate epitopes on the platelet membrane glycoprotein Ibalpha. Infect Immun 2004; 72: 6528-6537.
    CrossrefPubMedGoogle Scholar