Stoichiometric complex formation by proliferating cell nuclear antigen (PCNA) and its interacting protein: purification and crystallization of the DNA polymerase and PCNA monomer mutant complex fromPyrococcus furiosus
| Autor: | Daisuke Tsuchiya, Yoshizumi Ishino, Hirokazu Nishida, Shigeki Matsumiya, Kosuke Morikawa |
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| Rok vydání: | 2006 |
| Předmět: |
DNA clamp
biology DNA polymerase Biophysics DNA-Directed DNA Polymerase Processivity Crystallography X-Ray Condensed Matter Physics biology.organism_classification Biochemistry DNA polymerase delta Proliferating cell nuclear antigen Pyrococcus furiosus chemistry.chemical_compound chemistry Crystallization Communications Structural Biology Proliferating Cell Nuclear Antigen Protein purification Genetics biology.protein Crystallization DNA |
| Zdroj: | Acta Crystallographica Section F Structural Biology and Crystallization Communications. 62:253-256 |
| ISSN: | 1744-3091 |
| DOI: | 10.1107/s1744309106004362 |
| Popis: | Replicative DNA polymerase interacts with processivity factors, the beta-subunit of DNA polymerase III or proliferating cell nuclear antigen (PCNA), in order to function with a long template DNA. The archaeal replicative DNA polymerase from Pyrococcus furiosus interacts with PCNA via its PCNA-interacting protein (PIP) motif at the C-terminus. The PCNA homotrimeric ring contains one PIP interacting site on each monomer and since the ring can accommodate up to three molecules simultaneously, formation of a stable stoichiometric complex of PCNA with its interacting protein has been difficult to control in vitro. A stable complex of the DNA polymerase with PCNA, using a PCNA monomer mutant, has been purified and crystallized. The best ordered crystal diffracted to 3.0 A resolution using synchrotron radiation. The crystals belong to space group P2(1)2(1)2, with unit-cell parameters a = 225.3, b = 123.3, c = 91.3 A. |
| Databáze: | OpenAIRE |
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