1evw: Difference between revisions
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[[Image:1evw.gif|left|200px]] | [[Image:1evw.gif|left|200px]] | ||
'''L116A MUTANT OF THE HOMING ENDONUCLEASE I-PPOI COMPLEXED TO HOMING SITE DNA.''' | {{Structure | ||
|PDB= 1evw |SIZE=350|CAPTION= <scene name='initialview01'>1evw</scene>, resolution 3.1Å | |||
|SITE= | |||
|LIGAND= <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene> and <scene name='pdbligand=MG:MAGNESIUM ION'>MG</scene> | |||
|ACTIVITY= | |||
|GENE= | |||
}} | |||
'''L116A MUTANT OF THE HOMING ENDONUCLEASE I-PPOI COMPLEXED TO HOMING SITE DNA.''' | |||
==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
1EVW is a [ | 1EVW is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Physarum_polycephalum Physarum polycephalum]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1EVW OCA]. | ||
==Reference== | ==Reference== | ||
Conformational changes and cleavage by the homing endonuclease I-PpoI: a critical role for a leucine residue in the active site., Galburt EA, Chadsey MS, Jurica MS, Chevalier BS, Erho D, Tang W, Monnat RJ Jr, Stoddard BL, J Mol Biol. 2000 Jul 21;300(4):877-87. PMID:[http:// | Conformational changes and cleavage by the homing endonuclease I-PpoI: a critical role for a leucine residue in the active site., Galburt EA, Chadsey MS, Jurica MS, Chevalier BS, Erho D, Tang W, Monnat RJ Jr, Stoddard BL, J Mol Biol. 2000 Jul 21;300(4):877-87. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/10891275 10891275] | ||
[[Category: Physarum polycephalum]] | [[Category: Physarum polycephalum]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
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[[Category: dna binding b-sheets; c-terminal exchanged dimer interface; bent dna]] | [[Category: dna binding b-sheets; c-terminal exchanged dimer interface; bent dna]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 11:00:59 2008'' | ||
Revision as of 12:01, 20 March 2008
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| Coordinates: | save as pdb, mmCIF, xml | ||||||
L116A MUTANT OF THE HOMING ENDONUCLEASE I-PPOI COMPLEXED TO HOMING SITE DNA.
OverviewOverview
The homing endonuclease I-PpoI severely bends its DNA target, resulting in significant deformations of the minor and major groove near the scissile phosphate groups. To study the role of conformational changes within the protein catalyst and the DNA substrate, we have determined the structure of the enzyme in the absence of bound DNA, performed gel retardation analyses of DNA binding and bending, and have mutagenized a leucine residue that contacts an adenine nucleotide at the site of cleavage. The structure of the L116A/DNA complex has been determined and the effects of the mutation on affinity and catalysis have been measured. The wild-type protein displays a rigid-body rotation of its individual subunits upon DNA binding. Homing site DNA is not detectably bent in the absence of protein, but is sharply bent in both the wild-type and L116A complexes. These results indicate that binding involves a large distortion of the DNA and a smaller change in protein conformation. Leucine 116 is critical for binding and catalysis: it appears to be important for forming a well-ordered protein-DNA complex at the cleavage site, for maximal deformation of the DNA, and for desolvation of the nucleotide bases that are partially unstacked in the enzyme complex.
About this StructureAbout this Structure
1EVW is a Single protein structure of sequence from Physarum polycephalum. Full crystallographic information is available from OCA.
ReferenceReference
Conformational changes and cleavage by the homing endonuclease I-PpoI: a critical role for a leucine residue in the active site., Galburt EA, Chadsey MS, Jurica MS, Chevalier BS, Erho D, Tang W, Monnat RJ Jr, Stoddard BL, J Mol Biol. 2000 Jul 21;300(4):877-87. PMID:10891275
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