Purification, crystallization, X-ray diffraction analysis and phasing of an engineered single-chainPvuII restriction endonuclease
| Autor: | Elias Eliopoulos, Michael Kokkinidis, Dina Kotsifaki, Athanasios Hountas, Chrysi Meramveliotaki, Maria Androulaki |
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| Rok vydání: | 2007 |
| Předmět: |
Proteus vulgaris
Biophysics Protein Engineering Cleavage (embryo) Biochemistry law.invention chemistry.chemical_compound X-Ray Diffraction Structural Biology law Escherichia coli Genetics Molecular replacement Deoxyribonucleases Type II Site-Specific biology Protein engineering Condensed Matter Physics biology.organism_classification Recombinant Proteins Crystallography Restriction enzyme chemistry Crystallization Communications Recombinant DNA Crystallization Linker DNA |
| Zdroj: | Acta Crystallographica Section F Structural Biology and Crystallization Communications. 63:836-838 |
| ISSN: | 1744-3091 |
| DOI: | 10.1107/s1744309107040377 |
| Popis: | The restriction endonuclease PvuII from Proteus vulgaris has been converted from its wild-type homodimeric form into the enzymatically active single-chain variant scPvuII by tandemly joining the two subunits through the peptide linker Gly-Ser-Gly-Gly. scPvuII, which is suitable for the development of programmed restriction endonucleases for highly specific DNA cleavage, was purified and crystallized. The crystals diffract to a resolution of 2.35 A and belong to space group P4(2), with unit-cell parameters a = b = 101.92, c = 100.28 A and two molecules per asymmetric unit. Phasing was successfully performed by molecular replacement. |
| Databáze: | OpenAIRE |
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