1g9z

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{{STRUCTURE_1g9z| PDB=1g9z | SCENE= }}
{{STRUCTURE_1g9z| PDB=1g9z | SCENE= }}
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'''LAGLIDADG HOMING ENDONUCLEASE I-CREI / DNA PRODUCT COMPLEX WITH MAGNESIUM'''
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===LAGLIDADG HOMING ENDONUCLEASE I-CREI / DNA PRODUCT COMPLEX WITH MAGNESIUM===
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==Overview==
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Homing endonucleases, like restriction enzymes, cleave double-stranded DNA at specific target sites. The cleavage mechanism(s) utilized by LAGLIDADG endonucleases have been difficult to elucidate; their active sites are divergent, and only one low resolution cocrystal structure has been determined. Here we report two high resolution structures of the dimeric I-CreI homing endonuclease bound to DNA: a substrate complex with calcium and a product complex with magnesium. The bound metals in both complexes are verified by manganese anomalous difference maps. The active sites are positioned close together to facilitate cleavage across the DNA minor groove; each contains one metal ion bound between a conserved aspartate (Asp 20) and a single scissile phosphate. A third metal ion bridges the two active sites. This divalent cation is bound between aspartate residues from the active site of each subunit and is in simultaneous contact with the scissile phosphates of both DNA strands. A metal-bound water molecule acts as the nucleophile and is part of an extensive network of ordered water molecules that are positioned by enzyme side chains. These structures illustrate a unique variant of a two-metal endonuclease mechanism is employed by the highly divergent LAGLIDADG enzyme family.
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{{ABSTRACT_PUBMED_11276249}}
==About this Structure==
==About this Structure==
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[[Category: Laglidadg]]
[[Category: Laglidadg]]
[[Category: Nuclease mechanism]]
[[Category: Nuclease mechanism]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri May 2 17:19:56 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Tue Jul 1 04:58:12 2008''

Revision as of 01:58, 1 July 2008

Image:1g9z.png

Template:STRUCTURE 1g9z

LAGLIDADG HOMING ENDONUCLEASE I-CREI / DNA PRODUCT COMPLEX WITH MAGNESIUM

Template:ABSTRACT PUBMED 11276249

About this Structure

1G9Z is a Single protein structure of sequence from Chlamydomonas reinhardtii. Full crystallographic information is available from OCA.

Reference

The homing endonuclease I-CreI uses three metals, one of which is shared between the two active sites., Chevalier BS, Monnat RJ Jr, Stoddard BL, Nat Struct Biol. 2001 Apr;8(4):312-6. PMID:11276249

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