5OL1

Crystal structure of an inactivated Ssp SICLOPPS intein with a CAFHPQ extein

Summary for 5OL1

• Entry DOI: 10.2210/pdb5ol1/pdb
• Descriptor: DNA polymerase III subunit alpha,DNA polymerase III subunit alpha, 1,2-ETHANEDIOL, BETA-MERCAPTOETHANOL, ... (6 entities in total)
• Functional Keywords: intein, extein, siclopps, cyclic peptide, splicing
• Biological source: Synechocystis sp. (strain PCC 6803 / Kazusa); More
• Cellular location: Cytoplasm : P74750
• Total number of polymer chains: 1
• Total formula weight: 18934.17

Authors

Kick, L.M.,Schneider, S. (deposition date: 2017-07-26, release date: 2017-09-27, Last modification date: 2024-01-17)

Primary citation

Kick, L.M.,Harteis, S.,Koch, M.F.,Schneider, S.
Mechanistic Insights into Cyclic Peptide Generation by DnaE Split-Inteins through Quantitative and Structural Investigation.
Chembiochem, 18:2242-2246, 2017

PubMed Abstract: Inteins carry out protein-splicing reactions, which are used in protein chemistry, protein engineering and biotechnological applications. Rearrangement of the order of the domains in split-inteins results in head-to-tail cyclisation of the target sequence, which can be used for genetic encoding and expression of libraries of cyclic peptides (CPs). The efficiency of the splicing reaction depends on the target sequence. Here we used mass spectrometry to assess in vivo cyclic peptide formation from different hexameric target sequences by the DnaE split-inteins from Synechocystis sp. and Nostoc punctiforme, revealing a strong impact of the target sequence and of the intein on the intracellular peptide concentration. Furthermore, we determined the crystal structures of their pre-splicing complexes, which allowed us to identify F-block Asp17 as crucial for the DnaE-mediated splicing reaction.

PubMed: 28914478
DOI: 10.1002/cbic.201700503

Experimental method

X-RAY DIFFRACTION (1.751 Å)