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Sortase-click strategy for defined protein conjugation on a heptavalent cyclodextrin scaffold
Multivalent proteins or protein dendrimers are useful for clinical and biotechnological applications. However, assembly of chemically defined protein dendrimers is a challenging endeavor. In the past, majority of protein dendrimers have been developed on branched lysine scaffolds and are usually lim...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6534326/ https://www.ncbi.nlm.nih.gov/pubmed/31125361 http://dx.doi.org/10.1371/journal.pone.0217369 |
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author | Singh, Shikha Gupta, Kanchan Shukla, Shagun Sampathkumar, Srinivasa-Gopalan Roy, Rajendra P. |
author_facet | Singh, Shikha Gupta, Kanchan Shukla, Shagun Sampathkumar, Srinivasa-Gopalan Roy, Rajendra P. |
author_sort | Singh, Shikha |
collection | PubMed |
description | Multivalent proteins or protein dendrimers are useful for clinical and biotechnological applications. However, assembly of chemically defined protein dendrimers is a challenging endeavor. In the past, majority of protein dendrimers have been developed on branched lysine scaffolds and are usually limited to a valency of two to four. The naturally occurring cyclodextrin (CD) scaffold composed of 6–8 glucose units offers the possibility of expanding the valency. Here we have adapted a chemoenzymatic-click strategy for displaying heptavalent peptides and large proteins on the β-cyclodextrin (β-CD) scaffold. We demonstrate that recombinant proteins (engineered with a LPXTG pentapeptide motif at the carboxy terminus), labeled with an alkyne moiety by sortase-mediated ligation, can be easily clicked on to the azide-derivatized β-cyclodextrin through the Huisgen cycloaddition reaction yielding a well-defined heptavalent display of proteins. |
format | Online Article Text |
id | pubmed-6534326 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-65343262019-06-05 Sortase-click strategy for defined protein conjugation on a heptavalent cyclodextrin scaffold Singh, Shikha Gupta, Kanchan Shukla, Shagun Sampathkumar, Srinivasa-Gopalan Roy, Rajendra P. PLoS One Research Article Multivalent proteins or protein dendrimers are useful for clinical and biotechnological applications. However, assembly of chemically defined protein dendrimers is a challenging endeavor. In the past, majority of protein dendrimers have been developed on branched lysine scaffolds and are usually limited to a valency of two to four. The naturally occurring cyclodextrin (CD) scaffold composed of 6–8 glucose units offers the possibility of expanding the valency. Here we have adapted a chemoenzymatic-click strategy for displaying heptavalent peptides and large proteins on the β-cyclodextrin (β-CD) scaffold. We demonstrate that recombinant proteins (engineered with a LPXTG pentapeptide motif at the carboxy terminus), labeled with an alkyne moiety by sortase-mediated ligation, can be easily clicked on to the azide-derivatized β-cyclodextrin through the Huisgen cycloaddition reaction yielding a well-defined heptavalent display of proteins. Public Library of Science 2019-05-24 /pmc/articles/PMC6534326/ /pubmed/31125361 http://dx.doi.org/10.1371/journal.pone.0217369 Text en © 2019 Singh et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Singh, Shikha Gupta, Kanchan Shukla, Shagun Sampathkumar, Srinivasa-Gopalan Roy, Rajendra P. Sortase-click strategy for defined protein conjugation on a heptavalent cyclodextrin scaffold |
title | Sortase-click strategy for defined protein conjugation on a heptavalent cyclodextrin scaffold |
title_full | Sortase-click strategy for defined protein conjugation on a heptavalent cyclodextrin scaffold |
title_fullStr | Sortase-click strategy for defined protein conjugation on a heptavalent cyclodextrin scaffold |
title_full_unstemmed | Sortase-click strategy for defined protein conjugation on a heptavalent cyclodextrin scaffold |
title_short | Sortase-click strategy for defined protein conjugation on a heptavalent cyclodextrin scaffold |
title_sort | sortase-click strategy for defined protein conjugation on a heptavalent cyclodextrin scaffold |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6534326/ https://www.ncbi.nlm.nih.gov/pubmed/31125361 http://dx.doi.org/10.1371/journal.pone.0217369 |
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