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Publication Abstract from PubMed

Protein splicing in trans by split inteins has become a useful tool for protein engineering in vivo and in vitro. Inteins require Cys, Ser, or Thr at the first residue of C-terminal flanking sequence because a thiol or hydroxyl group in the side-chains is a nucleophile indispensable for trans-esterification step during protein splicing. Newly identified distinct sequences with homology to Hedgehog/Intein (Hint) superfamily, called Bacterial Intein-Like (BIL) domains, often do not have Cys, Ser, or Thr as the obligatory nucleophilic residue found in inteins. We demonstrated that BIL domains from Clostridium thermocellum (Cth) are proficient of protein splicing without any sequence changes. We determined the first solution NMR structure of a BIL domain, CthBIL4, to guide engineering of split BIL domains for protein ligation. The newly engineered split BIL domain could catalyze protein ligation by trans-splicing. Protein ligation without any nucleophilic residues of Cys, Ser, and Thr could alleviate junction sequence requirements for protein trans-splicing imposed by split inteins and could broaden applications of protein ligation by protein trans-splicing. This article is protected by copyright. All rights reserved.

Structural basis for Protein Trans-Splicing by a Bacterial Intein-Like domain: Protein Ligation without Nucleophilic side-chains.,Aranko AS, Oeemig JS, Iwai H FEBS J. 2013 Apr 27. doi: 10.1111/febs.12307. PMID:23621571^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.