Structure and Function Characterization of the a1a2 Motifs of Streptococcus pyogenes M Protein in Human Plasminogen Binding

Adam J.H. Quek, Blake A. Mazzitelli, Guojie Wu, Eleanor W.W. Leung, Tom T. Caradoc-Davies, Gordon J. Lloyd, Devadharshini Jeevarajah, Paul J. Conroy, Martina Sanderson-Smith, Yue Yuan, Yetunde A. Ayinuola, Francis J. Castellino, James C. Whisstock, Ruby H.P. Law

Research output: Contribution to journalArticleResearchpeer-review

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 languageEnglish
Number of pages10
JournalJournal of Molecular Biology
DOIs
Publication statusAccepted/In press - 8 Jul 2019

Keywords

  • a1a2 repeat
  • lysine-binding site
  • PAM
  • plasminogen kringle 2
  • RH motif

Cite this

@article{41016503b2554149a279e72a7b4e8144,
title = "Structure and Function Characterization of the a1a2 Motifs of Streptococcus pyogenes M Protein in Human Plasminogen Binding",
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-{\AA} 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.",
keywords = "a1a2 repeat, lysine-binding site, PAM, plasminogen kringle 2, RH motif",
author = "Quek, {Adam J.H.} and Mazzitelli, {Blake A.} and Guojie Wu and Leung, {Eleanor W.W.} and Caradoc-Davies, {Tom T.} and Lloyd, {Gordon J.} and Devadharshini Jeevarajah and Conroy, {Paul J.} and Martina Sanderson-Smith and Yue Yuan and Ayinuola, {Yetunde A.} and Castellino, {Francis J.} and Whisstock, {James C.} and Law, {Ruby H.P.}",
year = "2019",
month = "7",
day = "8",
doi = "10.1016/j.jmb.2019.07.003",
language = "English",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Elsevier",

}

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, 08.07.2019.

Research output: Contribution to journalArticleResearchpeer-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 - 2019/7/8

Y1 - 2019/7/8

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

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U2 - 10.1016/j.jmb.2019.07.003

DO - 10.1016/j.jmb.2019.07.003

M3 - Article

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

ER -