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

David M P De Oliveira, Ruby H P Law, Diane Ly, Simon M Cook, Adam J Quek, Jason D McArthur, James C Whisstock, Martina L Sanderson-Smith

Research output: Contribution to journalArticleResearchpeer-review

6 Citations (Scopus)


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 epsilon-aminocaproic acid (epsilonACA), whereas PAM-GII-Plg interactions were shown to be weakened but not inhibited in the presence of 400 mM epsilonACA. In contrast, binding to the KR1-3 domains of GII-Plg (angiostatin) by PAM was completely inhibited in the presence 5 mM epsilonACA. 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.
Original languageEnglish
Pages (from-to)3960 - 3968
Number of pages9
Issue number25
Publication statusPublished - 2015

Cite this