Structure determination of the O-methyltransferase NovP using the 'free lunch algorithm' as implemented in SHELXE
NovP is an S-adenosyl-l-methionine-dependent O-methyltransferase from Streptomyces spheroides (subunit MW = 29 967 Da). Recombinant N-terminally His-tagged NovP crystallizes in space group P2, with approximate unit-cell parameters a = 51.81, b = 46.04, c = 61.22 Å, β = 105.0°, giving a solvent conte...
| Autores: | , , , |
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| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2007 |
| País: | España |
| Institución: | Consejo Superior de Investigaciones Científicas (CSIC) |
| Repositorio: | DIGITAL.CSIC. Repositorio Institucional del CSIC |
| OAI Identifier: | oai:digital.csic.es:10261/113906 |
| Acceso en línea: | http://hdl.handle.net/10261/113906 |
| Access Level: | acceso abierto |
| Palabra clave: | Antibiotic biosynthesis Free lunch algorithm SHELXE NovP O-methyltransferase Chain tracing |
| Sumario: | NovP is an S-adenosyl-l-methionine-dependent O-methyltransferase from Streptomyces spheroides (subunit MW = 29 967 Da). Recombinant N-terminally His-tagged NovP crystallizes in space group P2, with approximate unit-cell parameters a = 51.81, b = 46.04, c = 61.22 Å, β = 105.0°, giving a solvent content of 44% for a single copy of the His-tagged protomer per asymmetric unit. Native synchrotron data to a resolution of 1.35 Å were combined with three other native data sets collected at lower resolution (both in-house and at the synchrotron) for the sake of completeness and better scaling. Data to 2.45 Å resolution were subsequently recorded in-house from a single mercury derivative. Three partial mercury sites could be located with SHELXD, but the resulting phases had a mean error of about 81° and in our hands did not yield an interpretable map using standard automated software. Nevertheless, the structure of NovP could be solved by first tracing a small part of the structure by hand and then extrapolating within and beyond the experimental resolution limit using the 'free lunch algorithm' in SHELXE. The resulting phases have a mean phase error of 17° relative to a refined model. © International Union of Crystallography 2007. |
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