Cargando…
Structures of Streptococcus pyogenes class A sortase in complex with substrate and product mimics provide key details of target recognition
The cell wall is a critical extracellular barrier for bacteria and many other organisms. In bacteria, this structural layer consists of peptidoglycan, which maintains cell shape and structural integrity and provides a scaffold for displaying various protein factors. To attach proteins to the cell wa...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9520033/ https://www.ncbi.nlm.nih.gov/pubmed/36055407 http://dx.doi.org/10.1016/j.jbc.2022.102446 |
_version_ | 1784799532530270208 |
---|---|
author | Johnson, D. Alex Piper, Isabel M. Vogel, Brandon A. Jackson, Sophie N. Svendsen, Justin E. Kodama, Hanna M. Lee, Darren E. Lindblom, Katy M. McCarty, James Antos, John M. Amacher, Jeanine F. |
author_facet | Johnson, D. Alex Piper, Isabel M. Vogel, Brandon A. Jackson, Sophie N. Svendsen, Justin E. Kodama, Hanna M. Lee, Darren E. Lindblom, Katy M. McCarty, James Antos, John M. Amacher, Jeanine F. |
author_sort | Johnson, D. Alex |
collection | PubMed |
description | The cell wall is a critical extracellular barrier for bacteria and many other organisms. In bacteria, this structural layer consists of peptidoglycan, which maintains cell shape and structural integrity and provides a scaffold for displaying various protein factors. To attach proteins to the cell wall, Gram-positive bacteria utilize sortase enzymes, which are cysteine transpeptidases that recognize and cleave a specific sorting signal, followed by ligation of the sorting signal–containing protein to the peptidoglycan precursor lipid II (LII). This mechanism is the subject of considerable interest as a target for therapeutic intervention and as a tool for protein engineering, where sortases have enabled sortase-mediated ligation or sortagging strategies. Despite these uses, there remains an incomplete understanding of the stereochemistry of substrate recognition and ligation product formation. Here, we solved the first structures of sortase A from Streptococcus pyogenes bound to two substrate sequences, LPATA and LPATS. In addition, we synthesized a mimetic of the product of sortase-mediated ligation involving LII (LPAT-LII) and solved the complex structure in two ligand conformations. These structures were further used as the basis for molecular dynamics simulations to probe sortase A-ligand dynamics and to construct a model of the acyl–enzyme intermediate, thus providing a structural view of multiple key states in the catalytic mechanism. Overall, this structural information provides new insights into the recognition of the sortase substrate motif and LII ligation partner and will support the continued development of sortases for protein engineering applications. |
format | Online Article Text |
id | pubmed-9520033 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-95200332022-10-04 Structures of Streptococcus pyogenes class A sortase in complex with substrate and product mimics provide key details of target recognition Johnson, D. Alex Piper, Isabel M. Vogel, Brandon A. Jackson, Sophie N. Svendsen, Justin E. Kodama, Hanna M. Lee, Darren E. Lindblom, Katy M. McCarty, James Antos, John M. Amacher, Jeanine F. J Biol Chem Research Article The cell wall is a critical extracellular barrier for bacteria and many other organisms. In bacteria, this structural layer consists of peptidoglycan, which maintains cell shape and structural integrity and provides a scaffold for displaying various protein factors. To attach proteins to the cell wall, Gram-positive bacteria utilize sortase enzymes, which are cysteine transpeptidases that recognize and cleave a specific sorting signal, followed by ligation of the sorting signal–containing protein to the peptidoglycan precursor lipid II (LII). This mechanism is the subject of considerable interest as a target for therapeutic intervention and as a tool for protein engineering, where sortases have enabled sortase-mediated ligation or sortagging strategies. Despite these uses, there remains an incomplete understanding of the stereochemistry of substrate recognition and ligation product formation. Here, we solved the first structures of sortase A from Streptococcus pyogenes bound to two substrate sequences, LPATA and LPATS. In addition, we synthesized a mimetic of the product of sortase-mediated ligation involving LII (LPAT-LII) and solved the complex structure in two ligand conformations. These structures were further used as the basis for molecular dynamics simulations to probe sortase A-ligand dynamics and to construct a model of the acyl–enzyme intermediate, thus providing a structural view of multiple key states in the catalytic mechanism. Overall, this structural information provides new insights into the recognition of the sortase substrate motif and LII ligation partner and will support the continued development of sortases for protein engineering applications. American Society for Biochemistry and Molecular Biology 2022-08-31 /pmc/articles/PMC9520033/ /pubmed/36055407 http://dx.doi.org/10.1016/j.jbc.2022.102446 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Article Johnson, D. Alex Piper, Isabel M. Vogel, Brandon A. Jackson, Sophie N. Svendsen, Justin E. Kodama, Hanna M. Lee, Darren E. Lindblom, Katy M. McCarty, James Antos, John M. Amacher, Jeanine F. Structures of Streptococcus pyogenes class A sortase in complex with substrate and product mimics provide key details of target recognition |
title | Structures of Streptococcus pyogenes class A sortase in complex with substrate and product mimics provide key details of target recognition |
title_full | Structures of Streptococcus pyogenes class A sortase in complex with substrate and product mimics provide key details of target recognition |
title_fullStr | Structures of Streptococcus pyogenes class A sortase in complex with substrate and product mimics provide key details of target recognition |
title_full_unstemmed | Structures of Streptococcus pyogenes class A sortase in complex with substrate and product mimics provide key details of target recognition |
title_short | Structures of Streptococcus pyogenes class A sortase in complex with substrate and product mimics provide key details of target recognition |
title_sort | structures of streptococcus pyogenes class a sortase in complex with substrate and product mimics provide key details of target recognition |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9520033/ https://www.ncbi.nlm.nih.gov/pubmed/36055407 http://dx.doi.org/10.1016/j.jbc.2022.102446 |
work_keys_str_mv | AT johnsondalex structuresofstreptococcuspyogenesclassasortaseincomplexwithsubstrateandproductmimicsprovidekeydetailsoftargetrecognition AT piperisabelm structuresofstreptococcuspyogenesclassasortaseincomplexwithsubstrateandproductmimicsprovidekeydetailsoftargetrecognition AT vogelbrandona structuresofstreptococcuspyogenesclassasortaseincomplexwithsubstrateandproductmimicsprovidekeydetailsoftargetrecognition AT jacksonsophien structuresofstreptococcuspyogenesclassasortaseincomplexwithsubstrateandproductmimicsprovidekeydetailsoftargetrecognition AT svendsenjustine structuresofstreptococcuspyogenesclassasortaseincomplexwithsubstrateandproductmimicsprovidekeydetailsoftargetrecognition AT kodamahannam structuresofstreptococcuspyogenesclassasortaseincomplexwithsubstrateandproductmimicsprovidekeydetailsoftargetrecognition AT leedarrene structuresofstreptococcuspyogenesclassasortaseincomplexwithsubstrateandproductmimicsprovidekeydetailsoftargetrecognition AT lindblomkatym structuresofstreptococcuspyogenesclassasortaseincomplexwithsubstrateandproductmimicsprovidekeydetailsoftargetrecognition AT mccartyjames structuresofstreptococcuspyogenesclassasortaseincomplexwithsubstrateandproductmimicsprovidekeydetailsoftargetrecognition AT antosjohnm structuresofstreptococcuspyogenesclassasortaseincomplexwithsubstrateandproductmimicsprovidekeydetailsoftargetrecognition AT amacherjeaninef structuresofstreptococcuspyogenesclassasortaseincomplexwithsubstrateandproductmimicsprovidekeydetailsoftargetrecognition |