1MOW

E-DreI

Summary for 1MOW

• Entry DOI: 10.2210/pdb1mow/pdb
• Descriptor: 5'-D(*CP*CP*AP*AP*AP*CP*TP*GP*TP*CP*TP*CP*AP*AP*GP*TP*TP*CP*CP*GP*GP*CP*G)-3', 5'-D(*CP*GP*CP*CP*GP*GP*AP*AP*CP*TP*TP*GP*AP*GP*AP*CP*AP*GP*TP*TP*TP*GP*G)-3', chimera of homing endonuclease I-DmoI and DNA endonuclease I-CreI, ... (7 entities in total)
• Functional Keywords: laglidadg, homing, engineering, design, endonuclease, hydrolase-dna complex, hydrolase/dna
• Biological source: Desulfurococcus mobilis (,); More
• Cellular location: Plastid, chloroplast: p05725
• Total number of polymer chains: 12
• Total formula weight: 179359.22

Authors

Chevalier, B.S.,Kortemme, T.,Chadsey, M.S.,Baker, D.,Monnat Jr., R.J.,Stoddard, B.L. (deposition date: 2002-09-10, release date: 2002-11-29, Last modification date: 2024-02-14)

Primary citation

Chevalier, B.S.,Kortemme, T.,Chadsey, M.S.,Baker, D.,Monnat Jr., R.J.,Stoddard, B.L.
Design, Activity and Structure of a Highly Specific Artificial Endonuclease
Mol.Cell, 10:895-905, 2002

PubMed Abstract: We have generated an artificial highly specific endonuclease by fusing domains of homing endonucleases I-DmoI and I-CreI and creating a new 1400 A(2) protein interface between these domains. Protein engineering was accomplished by combining computational redesign and an in vivo protein-folding screen. The resulting enzyme, E-DreI (Engineered I-DmoI/I-CreI), binds a long chimeric DNA target site with nanomolar affinity, cleaving it precisely at a rate equivalent to its natural parents. The structure of an E-DreI/DNA complex demonstrates the accuracy of the protein interface redesign algorithm and reveals how catalytic function is maintained during the creation of the new endonuclease. These results indicate that it may be possible to generate novel highly specific DNA binding proteins from homing endonucleases.

PubMed: 12419232
DOI: 10.1016/S1097-2765(02)00690-1

Experimental method

X-RAY DIFFRACTION (2.4 Å)