Structure-based group A streptococcal vaccine design: Helical wheel homology predicts antibody cross-reactivity among streptococcal M protein–derived peptides

Group A streptococcus (Strep A) surface M protein, an α-helical coiled-coil dimer, is a vaccine target and a major determinant of streptococcal virulence. The sequence-variable N-terminal region of the M protein defines the M type and also contains epitopes that promote opsonophagocytic killing of streptococci. Recent reports have reported considerable cross-reactivity among different M types, suggesting the prospect of identifying cross-protective epitopes that would constitute a broadly protective multivalent vaccine against Strep A isolates. Here, we have used a combination of immunological assays, structural biology, and cheminformatics to construct a recombinant M protein–based vaccine that included six Strep A M peptides that were predicted to elicit antisera that would cross-react with an additional 15 nonvaccine M types of Strep A. Rabbit antisera against this recombinant vaccine cross-reacted with 10 of the 15 nonvaccine M peptides. Two of the five nonvaccine M peptides that did not cross-react shared high sequence identity (≥50%) with the vaccine peptides, implying that high sequence identity alone was insufficient for cross-reactivity among the M peptides. Additional structural analyses revealed that the sequence identity at corresponding polar helical-wheel heptad sites between vaccine and nonvaccine peptides accurately distinguishes cross-reactive from non–cross-reactive peptides. On the basis of these observations, we developed a scoring algorithm based on the sequence identity at polar heptad sites. When applied to all epidemiologically important M types, this algorithm should enable the selection of a minimal number of M peptide–based vaccine candidates that elicit broadly protective immunity against Strep A.

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Binding of the kringle-2 domain of human plasminogen to streptococcal PAM-type M-protein causes dissociation of PAM dimers

10.22541/au.163257642.25497842/v1 ◽

2021 ◽

Author(s):

Olawole Ayinuola ◽

Yetunde Ayinuola ◽

Cunjia Qiu ◽

Shaun Lee ◽

Victoria Ploplis ◽

...

Keyword(s):

Group A Streptococcus ◽

Tight Binding ◽

Coiled Coil ◽

M Protein ◽

Temperature Dependent ◽

Functional Monomers ◽

Molecular Weights ◽

Group A ◽

Human Plasminogen ◽

Kringle 2

M-protein (PAM) largely contributes to the pathogenesis of Pattern D Group A Streptococcus pyogenes (GAS). However, the mechanism of complex formation is unknown. In a system consisting of a Class II PAM from Pattern D GAS isolate NS88.2 (PAMNS88.2), with one K2hPg binding a-repeat in its A-domain, we employed biophysical techniques to analyze the mechanism of the K2hPg/PAMNS88.2 interaction. We show that apo-PAMNS88.2 is a coiled-coil homodimer (M.Wt. ~80 kDa) at 4°C - 25°C, and is monomeric (M.Wt. ~40 kDa) at 37°C, demonstrating a temperature-dependent dissociation of PAMNS88.2 over a narrow temperature range. PAMNS88.2 displayed a single tight binding site for K2hPg at 4°C, which progressively increased at 25°C through 37°C. We isolated the K2hPg/PAMNS88.2 complexes at 4°C, 25°C, and 37°C and found molecular weights of ~50 kDa at each temperature, corresponding to a 1:1 (m:m) K2hPg/PAMNS88.2 monomer complex. hPg activation experiments by streptokinase demonstrated that the hPg/PAMNS88.2 monomer complexes are fully functional. The data show that PAM dimers dissociate into functional monomers at physiological temperatures or when presented with the active hPg module (K2hPg) showing that PAM is a functional monomer at 37°C.

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Protective Studies with Group A Streptococcal M Protein Vaccine. III. Challenge of Volunteers after Systemic or Intranasal Immunization with Type 3 or Type 12 Group A Streptococcus

The Journal of Infectious Diseases ◽

10.1093/infdis/138.6.712 ◽

1978 ◽

Vol 138 (6) ◽

pp. 712-718 ◽

Cited By ~ 44

Author(s):

R. D'Alessandri ◽

G. Plotkin ◽

R. M. Kluge ◽

M. K. Wittner ◽

E. N. Fox ◽

...

Keyword(s):

Group A Streptococcus ◽

M Protein ◽

Protein Vaccine ◽

Intranasal Immunization ◽

Streptococcal M Protein ◽

Group A ◽

Type 3

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Streptococcal M protein: molecular design and biological behavior.

Clinical Microbiology Reviews ◽

10.1128/cmr.2.3.285 ◽

1989 ◽

Vol 2 (3) ◽

pp. 285-314 ◽

Cited By ~ 509

Author(s):

V A Fischetti

Keyword(s):

Antigenic Variation ◽

Molecular Design ◽

Group A Streptococcus ◽

Coiled Coil ◽

Surface Proteins ◽

M Protein ◽

Biological Behavior ◽

Streptococcal M Protein ◽

Mammalian Proteins ◽

Common Motif

M protein is a major virulence determinant for the group A streptococcus by virtue of its ability to allow the organism to resist phagocytosis. Common in eucaryotes, the fibrillar coiled-coil design for the M molecule may prove to be a common motif for surface proteins in gram-positive organisms. This type of structure offers the organism several distinct advantages, ranging from antigenic variation to multiple functional domains. The close resemblance of this molecular design to that of certain mammalian proteins could help explain on a molecular level the formation of epitopes responsible for serological cross-reactions between microbial and mammalian proteins. Many of the approaches described in the elucidation of the M-protein structure may be applied for characterizing similar molecules in other microbial systems.

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The Plasminogen-Binding Group A Streptococcal M Protein-Related Protein Prp Binds Plasminogen via Arginine and Histidine Residues

Journal of Bacteriology ◽

10.1128/jb.01218-06 ◽

2006 ◽

Vol 189 (4) ◽

pp. 1435-1440 ◽

Cited By ~ 48

Author(s):

Martina L. Sanderson-Smith ◽

Mark Dowton ◽

Marie Ranson ◽

Mark J. Walker

Keyword(s):

Binding Site ◽

Group A Streptococcus ◽

Invasive Disease ◽

Site Directed Mutagenesis ◽

M Protein ◽

Histidine Residues ◽

Streptococcal M Protein ◽

M Proteins ◽

Lysine Residues ◽

Group A

ABSTRACT The migration of the human pathogen Streptococcus pyogenes (group A streptococcus) from localized to deep tissue sites may result in severe invasive disease, and sequestration of the host zymogen plasminogen appears crucial for virulence. Here, we describe a novel plasminogen-binding M protein, the plasminogen-binding group A streptococcal M protein (PAM)-related protein (Prp). Prp is phylogenetically distinct from previously described plasminogen-binding M proteins of group A, C, and G streptococci. While competition experiments indicate that Prp binds plasminogen with a lower affinity than PAM (50% effective concentration = 0.34 μM), Prp nonetheless binds plasminogen with high affinity and at physiologically relevant concentrations of plasminogen (Kd = 7.8 nM). Site-directed mutagenesis of the putative plasminogen binding site indicates that unlike the majority of plasminogen receptors, Prp does not interact with plasminogen exclusively via lysine residues. Mutagenesis to alanine of lysine residues Lys96 and Lys101 reduced but did not abrogate plasminogen binding by Prp. Plasminogen binding was abolished only with the additional mutagenesis of Arg107 and His108 to alanine. Furthermore, mutagenesis of Arg107 and His108 abolished plasminogen binding by Prp despite the presence of Lys96 and Lys101 in the binding site. Thus, binding to plasminogen via arginine and histidine residues appears to be a conserved mechanism among plasminogen-binding M proteins.

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Immunological Relationship between the Class I Epitope of Streptococcal M Protein and Myosin

Infection and Immunity ◽

10.1128/iai.66.9.4418-4424.1998 ◽

1998 ◽

Vol 66 (9) ◽

pp. 4418-4424 ◽

Cited By ~ 27

Author(s):

Anthony Quinn ◽

Kent Ward ◽

Vincent A. Fischetti ◽

Mark Hemric ◽

Madeleine W. Cunningham

Keyword(s):

Coiled Coil ◽

Class I ◽

M Protein ◽

Enzyme Linked Immunosorbent Assay ◽

Acute Glomerulonephritis ◽

Human Antibody ◽

Streptococcal M Protein ◽

Skeletal Myosin ◽

Group A ◽

Immunological Relationship

ABSTRACT The class I epitope of streptococcal M protein is an epidemiological marker for acute rheumatic fever (ARF)-associated serotypes of group A streptococci and is recognized by anti-M protein monoclonal antibody (MAb) 10B6. Using MAb 10B6, we determined the relationship between the class I epitope of M protein and the α-helical coiled-coil protein myosin. MAb 10B6 reacted by enzyme-linked immunosorbent assay and Western blotting with human cardiac myosin and rabbit skeletal myosin and its heavy meromyosin (HMM) subfragment. Overlapping synthetic peptides of M5 protein were used to identify the region of M5 protein recognized by MAb 10B6. Two C repeat peptides (C2A and C3) containing the amino acid sequence KGLRRDLDASREAK reacted with MAb 10B6. Partial sequence identity, RRDL, was found in the HMM fragment of myosin, which reacted with MAb 10B6. However, not all peptides of M5 protein and myosin containing the RRDL sequence reacted with MAb 10B6. ARF sera and sera from uncomplicated pharyngitis (UNC) reacted with C repeat region peptides of M protein, while acute glomerulonephritis sera were not as reactive. Affinity-purified human antibody to peptide C3 reacted with myosin. The data demonstrate that the class I epitope of M protein is immunologically cross-reactive with myosin and the HMM subfragment, and antibodies to peptide C3 and myosin were present in ARF and UNC sera.

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Group A streptococcal M protein activates the NLRP3 inflammasome

Nature Microbiology ◽

10.1038/s41564-017-0005-6 ◽

2017 ◽

Vol 2 (10) ◽

pp. 1425-1434 ◽

Cited By ~ 27

Author(s):

J. Andrés Valderrama ◽

Angelica M. Riestra ◽

Nina J. Gao ◽

Christopher N. LaRock ◽

Naveen Gupta ◽

...

Keyword(s):

Nlrp3 Inflammasome ◽

M Protein ◽

Streptococcal M Protein ◽

Group A

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Induction of human gamma interferon by structurally defined polypeptide fragments of group A streptococcal M protein.

Infection and Immunity ◽

10.1128/iai.43.1.122-126.1984 ◽

1984 ◽

Vol 43 (1) ◽

pp. 122-126 ◽

Cited By ~ 4

Author(s):

D A Weigent ◽

E H Beachey ◽

T Huff ◽

J W Peterson ◽

G J Stanton ◽

...

Keyword(s):

Gamma Interferon ◽

M Protein ◽

Streptococcal M Protein ◽

Group A

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Inhibition of the alternative C3 convertase and classical C5 convertase of complement by group A streptococcal M protein.

Infection and Immunity ◽

10.1128/iai.58.8.2535-2541.1990 ◽

1990 ◽

Vol 58 (8) ◽

pp. 2535-2541 ◽

Cited By ~ 15

Author(s):

K Hong ◽

T Kinoshita ◽

J Takeda ◽

H Kozono ◽

P Pramoonjago ◽

...

Keyword(s):

M Protein ◽

Streptococcal M Protein ◽

Group A

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Host Pathways of Hemostasis that Regulate Group A Streptococcus pyogenes Pathogenicity

Current Drug Targets ◽

10.2174/1389450120666190926152914 ◽

2020 ◽

Vol 21 (2) ◽

pp. 193-201

Author(s):

Victoria A. Ploplis ◽

Francis J. Castellino

Keyword(s):

Streptococcus Pyogenes ◽

Inflammatory Responses ◽

Group A Streptococcus ◽

Protein A ◽

M Protein ◽

Severe Group ◽

Group A ◽

Flow Through ◽

Hallmark Feature ◽

Major Target

A hallmark feature of severe Group A Streptococcus pyogenes (GAS) infection is dysregulated hemostasis. Hemostasis is the primary pathway for regulating blood flow through events that contribute towards clot formation and its dissolution. However, a number of studies have identified components of hemostasis in regulating survival and dissemination of GAS. Several proteins have been identified on the surface of GAS and they serve to either facilitate invasion to host distal sites or regulate inflammatory responses to the pathogen. GAS M-protein, a surface-exposed virulence factor, appears to be a major target for interactions with host hemostasis proteins. These interactions mediate biochemical events both on the surface of GAS and in the solution when M-protein is released into the surrounding environment through shedding or regulated proteolytic processes that dictate the fate of this pathogen. A thorough understanding of the mechanisms associated with these interactions could lead to novel approaches for altering the course of GAS pathogenicity.

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