Structure-Informed Design of an Enzymatically Inactive Vaccine Component for Group A <italic toggle="yes">Streptococcus</italic>
ABSTRACT Streptococcus pyogenes (group A Streptococcus [GAS]) causes ~700 million human infections/year, resulting in >500,000 deaths. There is no commercial GAS vaccine available. The GAS surface protein arginine deiminase (ADI) protects mice against a lethal challenge. ADI is an enzyme that con...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2013-08-01
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| Series: | mBio |
| Online Access: | https://journals.asm.org/doi/10.1128/mBio.00509-13 |
| _version_ | 1819011826969477120 |
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| author | Anna Henningham Daniel J. Ericsson Karla Langer Lachlan W. Casey Blagojce Jovcevski G. Singh Chhatwal J. Andrew Aquilina Michael R. Batzloff Bostjan Kobe Mark J. Walker |
| author_facet | Anna Henningham Daniel J. Ericsson Karla Langer Lachlan W. Casey Blagojce Jovcevski G. Singh Chhatwal J. Andrew Aquilina Michael R. Batzloff Bostjan Kobe Mark J. Walker |
| author_sort | Anna Henningham |
| collection | DOAJ |
| description | ABSTRACT Streptococcus pyogenes (group A Streptococcus [GAS]) causes ~700 million human infections/year, resulting in >500,000 deaths. There is no commercial GAS vaccine available. The GAS surface protein arginine deiminase (ADI) protects mice against a lethal challenge. ADI is an enzyme that converts arginine to citrulline and ammonia. Administration of a GAS vaccine preparation containing wild-type ADI, a protein with inherent enzymatic activity, may present a safety risk. In an approach intended to maximize the vaccine safety of GAS ADI, X-ray crystallography and structural immunogenic epitope mapping were used to inform vaccine design. This study aimed to knock out ADI enzyme activity without disrupting the three-dimensional structure or the recognition of immunogenic epitopes. We determined the crystal structure of ADI at 2.5 Å resolution and used it to select a number of amino acid residues for mutagenesis to alanine (D166, E220, H275, D277, and C401). Each mutant protein displayed abrogated activity, and three of the mutant proteins (those with the D166A, H275A, and D277A mutations) possessed a secondary structure and oligomerization state equivalent to those of the wild type, produced high-titer antisera, and avoided disruption of B-cell epitopes of ADI. In addition, antisera raised against the D166A and D277A mutant proteins bound to the GAS cell surface. The inactivated D166A and D277A mutant ADIs are ideal for inclusion in a GAS vaccine preparation. There is no human ortholog of ADI, and we confirm that despite limited structural similarity in the active-site region to human peptidyl ADI 4 (PAD4), ADI does not functionally mimic PAD4 and antiserum raised against GAS ADI does not recognize human PAD4. IMPORTANCE We present an example of structural biology informing human vaccine design. We previously showed that the administration of the enzyme arginine deiminase (ADI) to mice protected the mice against infection with multiple GAS serotypes. In this study, we determined the structure of GAS ADI and used this information to improve the vaccine safety of GAS ADI. Catalytically inactive mutant forms of ADI retained structure, recognition by antisera, and immunogenic epitopes, rendering them ideal for inclusion in GAS vaccine preparations. This example of structural biology informing vaccine design may underpin the formulation of a safe and efficacious GAS vaccine. |
| first_indexed | 2024-12-21T01:34:20Z |
| format | Article |
| id | doaj.art-90669fba22484c2091bf8b72cdcee539 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-12-21T01:34:20Z |
| publishDate | 2013-08-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-90669fba22484c2091bf8b72cdcee5392022-12-21T19:20:17ZengAmerican Society for MicrobiologymBio2150-75112013-08-014410.1128/mBio.00509-13Structure-Informed Design of an Enzymatically Inactive Vaccine Component for Group A <italic toggle="yes">Streptococcus</italic>Anna Henningham0Daniel J. Ericsson1Karla Langer2Lachlan W. Casey3Blagojce Jovcevski4G. Singh Chhatwal5J. Andrew Aquilina6Michael R. Batzloff7Bostjan Kobe8Mark J. Walker9School of Chemistry and Molecular Biosciences, Australian Infectious Diseases Research Centre, University of Queensland, St. Lucia, Qld., AustraliaSchool of Chemistry and Molecular Biosciences, Australian Infectious Diseases Research Centre, University of Queensland, St. Lucia, Qld., AustraliaSchool of Chemistry and Molecular Biosciences, Australian Infectious Diseases Research Centre, University of Queensland, St. Lucia, Qld., AustraliaSchool of Chemistry and Molecular Biosciences, Australian Infectious Diseases Research Centre, University of Queensland, St. Lucia, Qld., AustraliaIllawarra Health and Medical Research Institute and School of Biological Sciences, University of Wollongong, Wollongong, NSW, AustraliaDepartment of Medical Microbiology, Helmholtz Centre for Infection Research, Braunschweig, GermanyIllawarra Health and Medical Research Institute and School of Biological Sciences, University of Wollongong, Wollongong, NSW, AustraliaInstitute for Glycomics, Griffith University, Southport, Qld., AustraliaSchool of Chemistry and Molecular Biosciences, Australian Infectious Diseases Research Centre, University of Queensland, St. Lucia, Qld., AustraliaSchool of Chemistry and Molecular Biosciences, Australian Infectious Diseases Research Centre, University of Queensland, St. Lucia, Qld., AustraliaABSTRACT Streptococcus pyogenes (group A Streptococcus [GAS]) causes ~700 million human infections/year, resulting in >500,000 deaths. There is no commercial GAS vaccine available. The GAS surface protein arginine deiminase (ADI) protects mice against a lethal challenge. ADI is an enzyme that converts arginine to citrulline and ammonia. Administration of a GAS vaccine preparation containing wild-type ADI, a protein with inherent enzymatic activity, may present a safety risk. In an approach intended to maximize the vaccine safety of GAS ADI, X-ray crystallography and structural immunogenic epitope mapping were used to inform vaccine design. This study aimed to knock out ADI enzyme activity without disrupting the three-dimensional structure or the recognition of immunogenic epitopes. We determined the crystal structure of ADI at 2.5 Å resolution and used it to select a number of amino acid residues for mutagenesis to alanine (D166, E220, H275, D277, and C401). Each mutant protein displayed abrogated activity, and three of the mutant proteins (those with the D166A, H275A, and D277A mutations) possessed a secondary structure and oligomerization state equivalent to those of the wild type, produced high-titer antisera, and avoided disruption of B-cell epitopes of ADI. In addition, antisera raised against the D166A and D277A mutant proteins bound to the GAS cell surface. The inactivated D166A and D277A mutant ADIs are ideal for inclusion in a GAS vaccine preparation. There is no human ortholog of ADI, and we confirm that despite limited structural similarity in the active-site region to human peptidyl ADI 4 (PAD4), ADI does not functionally mimic PAD4 and antiserum raised against GAS ADI does not recognize human PAD4. IMPORTANCE We present an example of structural biology informing human vaccine design. We previously showed that the administration of the enzyme arginine deiminase (ADI) to mice protected the mice against infection with multiple GAS serotypes. In this study, we determined the structure of GAS ADI and used this information to improve the vaccine safety of GAS ADI. Catalytically inactive mutant forms of ADI retained structure, recognition by antisera, and immunogenic epitopes, rendering them ideal for inclusion in GAS vaccine preparations. This example of structural biology informing vaccine design may underpin the formulation of a safe and efficacious GAS vaccine.https://journals.asm.org/doi/10.1128/mBio.00509-13 |
| spellingShingle | Anna Henningham Daniel J. Ericsson Karla Langer Lachlan W. Casey Blagojce Jovcevski G. Singh Chhatwal J. Andrew Aquilina Michael R. Batzloff Bostjan Kobe Mark J. Walker Structure-Informed Design of an Enzymatically Inactive Vaccine Component for Group A <italic toggle="yes">Streptococcus</italic> mBio |
| title | Structure-Informed Design of an Enzymatically Inactive Vaccine Component for Group A <italic toggle="yes">Streptococcus</italic> |
| title_full | Structure-Informed Design of an Enzymatically Inactive Vaccine Component for Group A <italic toggle="yes">Streptococcus</italic> |
| title_fullStr | Structure-Informed Design of an Enzymatically Inactive Vaccine Component for Group A <italic toggle="yes">Streptococcus</italic> |
| title_full_unstemmed | Structure-Informed Design of an Enzymatically Inactive Vaccine Component for Group A <italic toggle="yes">Streptococcus</italic> |
| title_short | Structure-Informed Design of an Enzymatically Inactive Vaccine Component for Group A <italic toggle="yes">Streptococcus</italic> |
| title_sort | structure informed design of an enzymatically inactive vaccine component for group a italic toggle yes streptococcus italic |
| url | https://journals.asm.org/doi/10.1128/mBio.00509-13 |
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