Sortase-Catalyzed Initiator Attachment Enables High Yield Growth of a Stealth Polymer from the C Terminus of a Protein

Yizhi Qi, Miriam Amiram, Weiping Gao, Dewey G. McCafferty, Ashutosh Chilkoti

Research output: Contribution to journalArticlepeer-review

49 Scopus citations

Abstract

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. A general method is developed for the high-yield synthesis of C-terminally site-specific and one-to-one stoichiometric protein/peptide-polymer conjugates. Sortase A from Staphylococcus aureus is used to site-specifically attach an initiator solely at the C-terminus of green fluorescent protein, followed by in situ growth of a stealth polymer, poly(oligo(ethylene glycol) methyl ether methacrylate) by atom transfer radical polymerization.

Original languageEnglish
Pages (from-to)1256-1260
Number of pages5
JournalMacromolecular Rapid Communications
Volume34
Issue number15
DOIs
StatePublished - 1 Aug 2013
Externally publishedYes

Keywords

  • atom transfer radical polymerization (ATRP)
  • biological application of polymers
  • grafting from
  • protein-polymer conjugates
  • sortase

Fingerprint

Dive into the research topics of 'Sortase-Catalyzed Initiator Attachment Enables High Yield Growth of a Stealth Polymer from the C Terminus of a Protein'. Together they form a unique fingerprint.

Cite this