Protein engineering through tandem transamidation.

TitleProtein engineering through tandem transamidation.
Publication TypeJournal Article
Year of Publication2019
AuthorsThompson, RE, Stevens, AJ, Muir, TW
JournalNat Chem
Volume11
Issue8
Pagination737-743
Date Published2019 Aug
ISSN1755-4349
Abstract

Semisynthetic proteins engineered to contain non-coded elements such as post-translational modifications (PTMs) represent a powerful class of tools for interrogating biological processes. Here, we introduce a one-pot, chemoenzymatic method that allows broad access to chemically modified proteins. The approach involves a tandem transamidation reaction cascade that integrates intein-mediated protein splicing with enzyme-mediated peptide ligation. We show that this approach can be used to introduce PTMs and biochemical probes into a range of proteins including Cas9 nuclease and the transcriptional regulator MeCP2, which causes Rett syndrome when mutated. The versatility of the approach is further illustrated through the chemical tailoring of histone proteins within a native chromatin setting. We expect our approach will extend the scope of semisynthesis in protein engineering.

DOI10.1038/s41557-019-0281-2
Alternate JournalNat Chem
PubMed ID31263208
PubMed Central IDPMC6711197
Grant ListP01 CA196539 / CA / NCI NIH HHS / United States
R37 GM086868 / GM / NIGMS NIH HHS / United States