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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...

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Autores principales: Singh, Shikha, Gupta, Kanchan, Shukla, Shagun, Sampathkumar, Srinivasa-Gopalan, Roy, Rajendra P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
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.
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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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