1m0d
From Proteopedia
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'''Crystal Structure of Bacteriophage T7 Endonuclease I with a Wild-Type Active Site and Bound Manganese Ions''' | '''Crystal Structure of Bacteriophage T7 Endonuclease I with a Wild-Type Active Site and Bound Manganese Ions''' | ||
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==Reference== | ==Reference== | ||
Metal ions bound at the active site of the junction-resolving enzyme T7 endonuclease I., Hadden JM, Declais AC, Phillips SE, Lilley DM, EMBO J. 2002 Jul 1;21(13):3505-15. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/12093751 12093751] | Metal ions bound at the active site of the junction-resolving enzyme T7 endonuclease I., Hadden JM, Declais AC, Phillips SE, Lilley DM, EMBO J. 2002 Jul 1;21(13):3505-15. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/12093751 12093751] | ||
| - | [[Category: Deoxyribonuclease IV (phage-T(4)-induced)]] | ||
[[Category: Enterobacteria phage t7]] | [[Category: Enterobacteria phage t7]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
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[[Category: Lilley, D M.]] | [[Category: Lilley, D M.]] | ||
[[Category: Phillips, S E.]] | [[Category: Phillips, S E.]] | ||
| - | [[Category: | + | [[Category: Composite active site]] |
| - | [[Category: | + | [[Category: Domain swapped]] |
| - | [[Category: | + | [[Category: Holliday junction resolvase]] |
| - | [[Category: | + | [[Category: Homodimer]] |
| - | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 00:28:47 2008'' | |
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Revision as of 21:28, 2 May 2008
Crystal Structure of Bacteriophage T7 Endonuclease I with a Wild-Type Active Site and Bound Manganese Ions
Overview
T7 endonuclease I is a nuclease that is selective for the structure of the four-way DNA junction. The active site is similar to those of a number of restriction enzymes. We have solved the crystal structure of endonuclease I with a wild-type active site. Diffusion of manganese ions into the crystal revealed two peaks of electron density per active site, defining two metal ion-binding sites. Site 1 is fully occupied, and the manganese ion is coordinated by the carboxylate groups of Asp55 and Glu65, and the main chain carbonyl of Thr66. Site 2 is partially occupied, and the metal ion has a single protein ligand, the remaining carboxylate oxygen atom of Asp55. Isothermal titration calorimetry showed the sequential exothermic binding of two manganese ions in solution, with dissociation constants of 0.58 +/- 0.019 and 14 +/- 1.5 mM. These results are consistent with a two metal ion mechanism for the cleavage reaction, in which the hydrolytic water molecule is contained in the first coordination sphere of the site 1-bound metal ion.
About this Structure
1M0D is a Single protein structure of sequence from Enterobacteria phage t7. Full crystallographic information is available from OCA.
Reference
Metal ions bound at the active site of the junction-resolving enzyme T7 endonuclease I., Hadden JM, Declais AC, Phillips SE, Lilley DM, EMBO J. 2002 Jul 1;21(13):3505-15. PMID:12093751 Page seeded by OCA on Sat May 3 00:28:47 2008
