Open AccessSortase-click strategy for defined protein conjugation on a heptavalent cyclodextrin scaffold
Author(s) -
Shikha Singh,
Kumkum Gupta,
Shagun Shukla,
Srinivasa-Gopalan Sampathkumar,
Rajendra P. Roy
Publication year - 2019
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0217369
Subject(s) - sortase , sortase a , scaffold , click chemistry , chemistry , cyclodextrin , dendrimer , pentapeptide repeat , combinatorial chemistry , moiety , biochemistry , peptide , stereochemistry , computer science , database , bacterial protein , gene
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.