1mow: Difference between revisions
New page: left|200px<br /><applet load="1mow" size="450" color="white" frame="true" align="right" spinBox="true" caption="1mow, resolution 2.40Å" /> '''E-DreI'''<br /> ==O... |
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[[Image:1mow.gif|left|200px]]<br /><applet load="1mow" size=" | [[Image:1mow.gif|left|200px]]<br /><applet load="1mow" size="350" color="white" frame="true" align="right" spinBox="true" | ||
caption="1mow, resolution 2.40Å" /> | caption="1mow, resolution 2.40Å" /> | ||
'''E-DreI'''<br /> | '''E-DreI'''<br /> | ||
==Overview== | ==Overview== | ||
We have generated an artificial highly specific endonuclease by fusing | 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. | ||
==About this Structure== | ==About this Structure== | ||
1MOW is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Desulfurococcus_mobilis_and_chlamydomonas_reinhardtii Desulfurococcus mobilis and chlamydomonas reinhardtii] with MG, SO4 and GOL as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http:// | 1MOW is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Desulfurococcus_mobilis_and_chlamydomonas_reinhardtii Desulfurococcus mobilis and chlamydomonas reinhardtii] with <scene name='pdbligand=MG:'>MG</scene>, <scene name='pdbligand=SO4:'>SO4</scene> and <scene name='pdbligand=GOL:'>GOL</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1MOW OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Baker, D.]] | [[Category: Baker, D.]] | ||
[[Category: Chadsey, M | [[Category: Chadsey, M S.]] | ||
[[Category: Chevalier, B | [[Category: Chevalier, B S.]] | ||
[[Category: Jr., R | [[Category: Jr., R J.Monnat.]] | ||
[[Category: Kortemme, T.]] | [[Category: Kortemme, T.]] | ||
[[Category: Stoddard, B | [[Category: Stoddard, B L.]] | ||
[[Category: GOL]] | [[Category: GOL]] | ||
[[Category: MG]] | [[Category: MG]] | ||
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[[Category: structure]] | [[Category: structure]] | ||
''Page seeded by [http:// | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:57:27 2008'' | ||
Revision as of 14:57, 21 February 2008
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E-DreI
OverviewOverview
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.
About this StructureAbout this Structure
1MOW is a Single protein structure of sequence from Desulfurococcus mobilis and chlamydomonas reinhardtii with , and as ligands. Full crystallographic information is available from OCA.
ReferenceReference
Design, activity, and structure of a highly specific artificial endonuclease., Chevalier BS, Kortemme T, Chadsey MS, Baker D, Monnat RJ, Stoddard BL, Mol Cell. 2002 Oct;10(4):895-905. PMID:12419232
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