2LWY: Solution Structure of Bacterial Intein-Like domain from Clostridium thermocellum

UNLABELLED: 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 the C-terminal flanking sequence because a thiol or hydroxyl group in the side chains is a nucleophile indispensable for the trans-esterification step during protein splicing. Newly-identified distinct sequences with homology to the hedgehog/intein 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 at 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. DATABASE: The resonance assignments and structure coordinates have been deposited in BMRB (18653) and RCSB (2LWY).
PDB ID: 2LWYDownload
MMDB ID: 109777
PDB Deposition Date: 2012/8/9
Updated in MMDB: 2013/07
Experimental Method:
solution nmr
Source Organism:
Similar Structures:
Biological Unit for 2LWY: monomeric; determined by author
Molecular Components in 2LWY
Label Count Molecule
Protein (1 molecule)
Bacterial Intein-like Domain
Molecule annotation
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Citing MMDB