2oxv: Difference between revisions
New page: left|200px<br /><applet load="2oxv" size="350" color="white" frame="true" align="right" spinBox="true" caption="2oxv, resolution 1.950Å" /> '''Structure of the A1... |
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==Overview== | ==Overview== | ||
Promiscuous mutant EcoRI endonucleases bind to the canonical site GAATTC | Promiscuous mutant EcoRI endonucleases bind to the canonical site GAATTC more tightly than does the wild-type endonuclease, yet cleave variant (EcoRI(*)) sites more rapidly than does wild-type. The crystal structure of the A138T promiscuous mutant homodimer in complex with a GAATTC site is nearly identical to that of the wild-type complex, except that the Thr138 side chains make packing interactions with bases in the 5'-flanking regions outside the recognition hexanucleotide while excluding two bound water molecules seen in the wild-type complex. Molecular dynamics simulations confirm exclusion of these waters. The structure and simulations suggest possible reasons why binding of the A138T protein to the GAATTC site has DeltaS degrees more favorable and DeltaH degrees less favorable than for wild-type endonuclease binding. The interactions of Thr138 with flanking bases may permit A138T, unlike wild-type enzyme, to form complexes with EcoRI(*) sites that structurally resemble the specific wild-type complex with GAATTC. | ||
==About this Structure== | ==About this Structure== | ||
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[[Category: Type II site-specific deoxyribonuclease]] | [[Category: Type II site-specific deoxyribonuclease]] | ||
[[Category: Jen-Jacobson, L.]] | [[Category: Jen-Jacobson, L.]] | ||
[[Category: Rosenberg, J | [[Category: Rosenberg, J M.]] | ||
[[Category: Sapienza, P | [[Category: Sapienza, P J.]] | ||
[[Category: ecori]] | [[Category: ecori]] | ||
[[Category: hydrolase/dna complex]] | [[Category: hydrolase/dna complex]] | ||
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[[Category: type ii restriction endonuclease]] | [[Category: type ii restriction endonuclease]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:23:45 2008'' | ||
Revision as of 19:23, 21 February 2008
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Structure of the A138T promiscuous mutant of the EcoRI restriction endonuclease bound to its cognate recognition site.
OverviewOverview
Promiscuous mutant EcoRI endonucleases bind to the canonical site GAATTC more tightly than does the wild-type endonuclease, yet cleave variant (EcoRI(*)) sites more rapidly than does wild-type. The crystal structure of the A138T promiscuous mutant homodimer in complex with a GAATTC site is nearly identical to that of the wild-type complex, except that the Thr138 side chains make packing interactions with bases in the 5'-flanking regions outside the recognition hexanucleotide while excluding two bound water molecules seen in the wild-type complex. Molecular dynamics simulations confirm exclusion of these waters. The structure and simulations suggest possible reasons why binding of the A138T protein to the GAATTC site has DeltaS degrees more favorable and DeltaH degrees less favorable than for wild-type endonuclease binding. The interactions of Thr138 with flanking bases may permit A138T, unlike wild-type enzyme, to form complexes with EcoRI(*) sites that structurally resemble the specific wild-type complex with GAATTC.
About this StructureAbout this Structure
2OXV is a Protein complex structure of sequences from Escherichia coli. Active as Type II site-specific deoxyribonuclease, with EC number 3.1.21.4 Full crystallographic information is available from OCA.
ReferenceReference
Structural and thermodynamic basis for enhanced DNA binding by a promiscuous mutant EcoRI endonuclease., Sapienza PJ, Rosenberg JM, Jen-Jacobson L, Structure. 2007 Nov;15(11):1368-82. PMID:17997963
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