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Preferential acquisition and activation of plasminogen glycoform II by PAM positive group A streptococcal isolates

Journal Article


Abstract


  • Plasminogen (Plg) circulates in the host as two predominant glycoforms. Glycoform I Plg (GI-Plg) contains glycosylation sites at Asn289 and Thr346, whereas glycoform II Plg (GII-Plg) is exclusively glycosylated at Thr346. Surface plasmon resonance experiments demonstrated that Plg binding group A streptococcal M protein (PAM) exhibits comparative equal affinity for GI- and GII-Plg in the “closed” conformation (for GII-Plg, KD = 27.4 nM; for GI-Plg, KD = 37.0 nM). When Plg was in the “open” conformation, PAM exhibited an 11-fold increase in affinity for GII-Plg (KD = 2.8 nM) compared with that for GI-Plg (KD = 33.2 nM). The interaction of PAM with Plg is believed to be mediated by lysine binding sites within kringle (KR) 2 of Plg. PAM–GI-Plg interactions were fully inhibited with 100 mM lysine analogue ε-aminocaproic acid (εACA), whereas PAM–GII-Plg interactions were shown to be weakened but not inhibited in the presence of 400 mM εACA. In contrast, binding to the KR1–3 domains of GII-Plg (angiostatin) by PAM was completely inhibited in the presence 5 mM εACA. Along with PAM, emm pattern D GAS isolates express a phenotypically distinct SK variant (type 2b SK) that requires Plg ligands such as PAM to activate Plg. Type 2b SK was able to generate an active site and activate GII-Plg at a rate significantly higher than that of GI-Plg when bound to PAM. Taken together, these data suggest that GAS selectively recruits and activates GII-Plg. Furthermore, we propose that the interaction between PAM and Plg may be partially mediated by a secondary binding site outside of KR2, affected by glycosylation at Asn289.

Publication Date


  • 2015

Citation


  • De Oliveira, D. M.P., Law, R. H. P., Ly, D., Cook, S. M., Quek, A. J., McArthur, J. D., Whisstock, J. C. & Sanderson-Smith, M. L. (2015). Preferential acquisition and activation of plasminogen glycoform II by PAM positive group A streptococcal isolates. Biochemistry, 54 (25), 3960-3968.

Scopus Eid


  • 2-s2.0-84933564452

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/3006

Has Global Citation Frequency


Number Of Pages


  • 8

Start Page


  • 3960

End Page


  • 3968

Volume


  • 54

Issue


  • 25

Place Of Publication


  • United States

Abstract


  • Plasminogen (Plg) circulates in the host as two predominant glycoforms. Glycoform I Plg (GI-Plg) contains glycosylation sites at Asn289 and Thr346, whereas glycoform II Plg (GII-Plg) is exclusively glycosylated at Thr346. Surface plasmon resonance experiments demonstrated that Plg binding group A streptococcal M protein (PAM) exhibits comparative equal affinity for GI- and GII-Plg in the “closed” conformation (for GII-Plg, KD = 27.4 nM; for GI-Plg, KD = 37.0 nM). When Plg was in the “open” conformation, PAM exhibited an 11-fold increase in affinity for GII-Plg (KD = 2.8 nM) compared with that for GI-Plg (KD = 33.2 nM). The interaction of PAM with Plg is believed to be mediated by lysine binding sites within kringle (KR) 2 of Plg. PAM–GI-Plg interactions were fully inhibited with 100 mM lysine analogue ε-aminocaproic acid (εACA), whereas PAM–GII-Plg interactions were shown to be weakened but not inhibited in the presence of 400 mM εACA. In contrast, binding to the KR1–3 domains of GII-Plg (angiostatin) by PAM was completely inhibited in the presence 5 mM εACA. Along with PAM, emm pattern D GAS isolates express a phenotypically distinct SK variant (type 2b SK) that requires Plg ligands such as PAM to activate Plg. Type 2b SK was able to generate an active site and activate GII-Plg at a rate significantly higher than that of GI-Plg when bound to PAM. Taken together, these data suggest that GAS selectively recruits and activates GII-Plg. Furthermore, we propose that the interaction between PAM and Plg may be partially mediated by a secondary binding site outside of KR2, affected by glycosylation at Asn289.

Publication Date


  • 2015

Citation


  • De Oliveira, D. M.P., Law, R. H. P., Ly, D., Cook, S. M., Quek, A. J., McArthur, J. D., Whisstock, J. C. & Sanderson-Smith, M. L. (2015). Preferential acquisition and activation of plasminogen glycoform II by PAM positive group A streptococcal isolates. Biochemistry, 54 (25), 3960-3968.

Scopus Eid


  • 2-s2.0-84933564452

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/3006

Has Global Citation Frequency


Number Of Pages


  • 8

Start Page


  • 3960

End Page


  • 3968

Volume


  • 54

Issue


  • 25

Place Of Publication


  • United States