5ESP: Crystal Structure Of Laglidadg Meganuclease I-panmi With Coordinated Calcium Ions

Citation:
Abstract
LAGLIDADG meganucleases are DNA cleaving enzymes used for genome engineering. While their cleavage specificity can be altered using several protein engineering and selection strategies, their overall targetability is limited by highly specific indirect recognition of the central four base pairs within their recognition sites. In order to examine the physical basis of indirect sequence recognition and to expand the number of such nucleases available for genome engineering, we have determined the target sites, DNA-bound structures, and central four cleavage fidelities of nine related enzymes. Subsequent crystallographic analyses of a meganuclease bound to two noncleavable target sites, each containing a single inactivating base pair substitution at its center, indicates that a localized slip of the mutated base pair causes a small change in the DNA backbone conformation that results in a loss of metal occupancy at one binding site, eliminating cleavage activity.
PDB ID: 5ESPDownload
MMDB ID: 137915
PDB Deposition Date: 2015/11/16
Updated in MMDB: 2017/09
Experimental Method:
x-ray diffraction
Resolution: 3  Å
Source Organism:
Podospora anserina S mat+
Similar Structures:
Biological Unit for 5ESP: trimeric; determined by author and by software (PISA)
Molecular Components in 5ESP
Label Count Molecule
Protein (1 molecule)
1
I-panmi
Molecule annotation
Nucleotides(2 molecules)
1
DNA (27-mer)
Molecule annotation
1
DNA (27-mer)
Molecule annotation
Chemicals (3 molecules)
1
2
2
1
* Click molecule labels to explore molecular sequence information.

Citing MMDB
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