Sortase-catalyzed initiator attachment enables high yield growth of a stealth polymer from the C terminus of a protein.
Journal Article (Journal Article)
Conventional methods for synthesizing protein/peptide-polymer conjugates, as a means to improve the pharmacological properties of therapeutic biomolecules, typically have drawbacks including low yield, non-trivial separation of conjugates from reactants, and lack of site- specificity, which results in heterogeneous products with significantly compromised bioactivity. To address these limitations, the use of sortase A from Staphylococcus aureus is demonstrated to site-specifically attach an initiator solely at the C-terminus of green fluorescent protein (GFP), followed by in situ growth of a stealth polymer, poly(oligo(ethylene glycol) methyl ether methacrylate) by atom transfer radical polymerization (ATRP). Sortase-catalyzed initiator attachment proceeds with high specificity and near-complete (≈95%) product conversion. Subsequent in situ ATRP in aqueous buffer produces 1:1 stoichiometric conjugates with >90% yield, low dispersity, and no denaturation of the protein. This approach introduces a simple and useful method for high yield synthesis of protein/peptide-polymer conjugates.
Full Text
Duke Authors
Cited Authors
- Qi, Y; Amiram, M; Gao, W; McCafferty, DG; Chilkoti, A
Published Date
- August 2013
Published In
Volume / Issue
- 34 / 15
Start / End Page
- 1256 - 1260
PubMed ID
- 23836349
Pubmed Central ID
- PMC3754797
Electronic International Standard Serial Number (EISSN)
- 1521-3927
International Standard Serial Number (ISSN)
- 1022-1336
Digital Object Identifier (DOI)
- 10.1002/marc.201300460
Language
- eng