Abstract
Plasminogen (Plg)-binding M protein (PAM) is a group A streptococcal cell surface receptor that is crucial for bacterial virulence. Previous studies revealed that, by binding to the kringle 2 (KR2) domain of host Plg, the pathogen attains a proteolytic microenvironment on the cell surface that facilitates its dissemination from the primary infection site. Each of the PAM molecules in their dimeric assembly consists of two Plg binding motifs (called the a1 and a2 repeats). To date, the molecular interactions between the a1 repeat and KR2 have been structurally characterized, whereas the role of the a2 repeat is less well defined. Here, we report the 1.7-Å x-ray crystal structure of KR2 in complex with a monomeric PAM peptide that contains both the a1 and a2 motifs. The structure reveals how the PAM peptide forms key interactions simultaneously with two KR2 via the high-affinity lysine isosteres within the a1a2 motifs. Further studies, through combined mutagenesis and functional characterization, show that a2 is a stronger KR2 binder than a1, suggesting that these two motifs may play discrete roles in mediating the final PAM-Plg assembly.
| Original language | English |
|---|---|
| Pages (from-to) | 3804-3813 |
| Number of pages | 10 |
| Journal | Journal of Molecular Biology |
| Volume | 431 |
| Issue number | 19 |
| DOIs | |
| Publication status | Published - 6 Sep 2019 |
Keywords
- a1a2 repeat
- lysine-binding site
- PAM
- plasminogen kringle 2
- RH motif
Cite this
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Structure and Function Characterization of the a1a2 Motifs of Streptococcus pyogenes M Protein in Human Plasminogen Binding. / Quek, Adam J.H.; Mazzitelli, Blake A.; Wu, Guojie; Leung, Eleanor W.W.; Caradoc-Davies, Tom T.; Lloyd, Gordon J.; Jeevarajah, Devadharshini; Conroy, Paul J.; Sanderson-Smith, Martina; Yuan, Yue; Ayinuola, Yetunde A.; Castellino, Francis J.; Whisstock, James C.; Law, Ruby H.P.
In: Journal of Molecular Biology, Vol. 431, No. 19, 06.09.2019, p. 3804-3813.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Structure and Function Characterization of the a1a2 Motifs of Streptococcus pyogenes M Protein in Human Plasminogen Binding
AU - Quek, Adam J.H.
AU - Mazzitelli, Blake A.
AU - Wu, Guojie
AU - Leung, Eleanor W.W.
AU - Caradoc-Davies, Tom T.
AU - Lloyd, Gordon J.
AU - Jeevarajah, Devadharshini
AU - Conroy, Paul J.
AU - Sanderson-Smith, Martina
AU - Yuan, Yue
AU - Ayinuola, Yetunde A.
AU - Castellino, Francis J.
AU - Whisstock, James C.
AU - Law, Ruby H.P.
PY - 2019/9/6
Y1 - 2019/9/6
N2 - Plasminogen (Plg)-binding M protein (PAM) is a group A streptococcal cell surface receptor that is crucial for bacterial virulence. Previous studies revealed that, by binding to the kringle 2 (KR2) domain of host Plg, the pathogen attains a proteolytic microenvironment on the cell surface that facilitates its dissemination from the primary infection site. Each of the PAM molecules in their dimeric assembly consists of two Plg binding motifs (called the a1 and a2 repeats). To date, the molecular interactions between the a1 repeat and KR2 have been structurally characterized, whereas the role of the a2 repeat is less well defined. Here, we report the 1.7-Å x-ray crystal structure of KR2 in complex with a monomeric PAM peptide that contains both the a1 and a2 motifs. The structure reveals how the PAM peptide forms key interactions simultaneously with two KR2 via the high-affinity lysine isosteres within the a1a2 motifs. Further studies, through combined mutagenesis and functional characterization, show that a2 is a stronger KR2 binder than a1, suggesting that these two motifs may play discrete roles in mediating the final PAM-Plg assembly.
AB - Plasminogen (Plg)-binding M protein (PAM) is a group A streptococcal cell surface receptor that is crucial for bacterial virulence. Previous studies revealed that, by binding to the kringle 2 (KR2) domain of host Plg, the pathogen attains a proteolytic microenvironment on the cell surface that facilitates its dissemination from the primary infection site. Each of the PAM molecules in their dimeric assembly consists of two Plg binding motifs (called the a1 and a2 repeats). To date, the molecular interactions between the a1 repeat and KR2 have been structurally characterized, whereas the role of the a2 repeat is less well defined. Here, we report the 1.7-Å x-ray crystal structure of KR2 in complex with a monomeric PAM peptide that contains both the a1 and a2 motifs. The structure reveals how the PAM peptide forms key interactions simultaneously with two KR2 via the high-affinity lysine isosteres within the a1a2 motifs. Further studies, through combined mutagenesis and functional characterization, show that a2 is a stronger KR2 binder than a1, suggesting that these two motifs may play discrete roles in mediating the final PAM-Plg assembly.
KW - a1a2 repeat
KW - lysine-binding site
KW - PAM
KW - plasminogen kringle 2
KW - RH motif
UR - http://www.scopus.com/inward/record.url?scp=85069214088&partnerID=8YFLogxK
U2 - 10.1016/j.jmb.2019.07.003
DO - 10.1016/j.jmb.2019.07.003
M3 - Article
C2 - 31295457
AN - SCOPUS:85069214088
VL - 431
SP - 3804
EP - 3813
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
SN - 0022-2836
IS - 19
ER -