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

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2 Citations (Scopus)

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 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
JournalBiochemistry
Volume54
Issue number25
DOIs
Publication statusPublished - 2015

Cite this

De Oliveira, David M P ; Law, Ruby H P ; Ly, Diane ; Cook, Simon M ; Quek, Adam J ; McArthur, Jason D ; Whisstock, James C ; Sanderson-Smith, Martina L. / Preferential acquisition and activation of plasminogen glycoform II by PAM positive group A streptococcal isolates. In: Biochemistry. 2015 ; Vol. 54, No. 25. pp. 3960 - 3968.
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title = "Preferential acquisition and activation of plasminogen glycoform II by PAM positive group A streptococcal isolates",
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 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.",
author = "{De Oliveira}, {David M P} and Law, {Ruby H P} and Diane Ly and Cook, {Simon M} and Quek, {Adam J} and McArthur, {Jason D} and Whisstock, {James C} and Sanderson-Smith, {Martina L}",
year = "2015",
doi = "10.1021/acs.biochem.5b00130",
language = "English",
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pages = "3960 -- 3968",
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Preferential acquisition and activation of plasminogen glycoform II by PAM positive group A streptococcal isolates. / De Oliveira, David M P; Law, Ruby H P; Ly, Diane; Cook, Simon M; Quek, Adam J; McArthur, Jason D; Whisstock, James C; Sanderson-Smith, Martina L.

In: Biochemistry, Vol. 54, No. 25, 2015, p. 3960 - 3968.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Preferential acquisition and activation of plasminogen glycoform II by PAM positive group A streptococcal isolates

AU - De Oliveira, David M P

AU - Law, Ruby H P

AU - Ly, Diane

AU - Cook, Simon M

AU - Quek, Adam J

AU - McArthur, Jason D

AU - Whisstock, James C

AU - Sanderson-Smith, Martina L

PY - 2015

Y1 - 2015

N2 - 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.

AB - 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.

UR - http://pubs.acs.org/doi/pdf/10.1021/acs.biochem.5b00130

U2 - 10.1021/acs.biochem.5b00130

DO - 10.1021/acs.biochem.5b00130

M3 - Article

VL - 54

SP - 3960

EP - 3968

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 25

ER -