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2pfo
From Proteopedia
| 2pfo, resolution 2.00Å () | |||||||||
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| Ligands: | , , , , | ||||||||
| Gene: | POLL (Homo sapiens) | ||||||||
| Related: | 2pfn, 2pfp, 2pfq | ||||||||
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| Resources: | FirstGlance, OCA, PDBsum, RCSB | ||||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||||
DNA Polymerase lambda in complex with DNA and dUPNPP
The incorporation of dNMPs into DNA by polymerases involves a phosphoryl transfer reaction hypothesized to require two divalent metal ions. Here we investigate this hypothesis using as a model human DNA polymerase lambda (Pol lambda), an enzyme suggested to be activated in vivo by manganese. We report the crystal structures of four complexes of human Pol lambda. In a 1.9 A structure of Pol lambda containing a 3'-OH and the non-hydrolyzable analog dUpnpp, a non-catalytic Na+ ion occupies the site for metal A and the ribose of the primer-terminal nucleotide is found in a conformation that positions the acceptor 3'-OH out of line with the alpha-phosphate and the bridging oxygen of the pyrophosphate leaving group. Soaking this crystal in MnCl2 yielded a 2.0 A structure with Mn2+ occupying the site for metal A. In the presence of Mn2+, the conformation of the ribose is C3'-endo and the 3'-oxygen is in line with the leaving oxygen, at a distance from the phosphorus atom of the alpha-phosphate (3.69 A) consistent with and supporting a catalytic mechanism involving two divalent metal ions. Finally, soaking with MnCl2 converted a pre-catalytic Pol lambda/Na+ complex with unreacted dCTP in the active site into a product complex via catalysis in the crystal. These data provide pre- and post-transition state information and outline in a single crystal the pathway for the phosphoryl transfer reaction carried out by DNA polymerases.
Role of the catalytic metal during polymerization by DNA polymerase lambda., Garcia-Diaz M, Bebenek K, Krahn JM, Pedersen LC, Kunkel TA, DNA Repair (Amst). 2007 Sep 1;6(9):1333-40. Epub 2007 May 1. PMID:17475573
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
About this Structure
2PFO is a 4 chains structure of sequences from Homo sapiens. Full crystallographic information is available from OCA.
Reference
- Garcia-Diaz M, Bebenek K, Krahn JM, Pedersen LC, Kunkel TA. Role of the catalytic metal during polymerization by DNA polymerase lambda. DNA Repair (Amst). 2007 Sep 1;6(9):1333-40. Epub 2007 May 1. PMID:17475573 doi:10.1016/j.dnarep.2007.03.005
Page seeded by OCA on Tue Feb 17 11:53:30 2009
