Structure of the cyanobactin oxidase ThcOx from Cyanothece sp. PCC 7425, the first structure to be solved at Diamond Light Source beamline I23 by means of S-SAD.Report as inadecuate




Structure of the cyanobactin oxidase ThcOx from Cyanothece sp. PCC 7425, the first structure to be solved at Diamond Light Source beamline I23 by means of S-SAD. - Download this document for free, or read online. Document in PDF available to download.

Reference: Bent, AF, Mann, G, Houssen, WE et al., (2017). Structure of the cyanobactin oxidase ThcOx from Cyanothece sp. PCC 7425, the first structure to be solved at Diamond Light Source beamline I23 by means of S-SAD. Acta Crystallographica Section D: Structural Biology, 72 (Pt 11), 1174-1180.Citable link to this page:

 

Structure of the cyanobactin oxidase ThcOx from Cyanothece sp. PCC 7425, the first structure to be solved at Diamond Light Source beamline I23 by means of S-SAD.

Abstract: Determination of protein crystal structures requires that the phases are derived independently of the observed measurement of diffraction intensities. Many techniques have been developed to obtain phases, including heavy-atom substitution, molecular replacement and substitution during protein expression of the amino acid methionine with selenomethionine. Although the use of selenium-containing methionine has transformed the experimental determination of phases it is not always possible, either because the variant protein cannot be produced or does not crystallize. Phasing of structures by measuring the anomalous diffraction from S atoms could in theory be almost universal since almost all proteins contain methionine or cysteine. Indeed, many structures have been solved by the so-called native sulfur single-wavelength anomalous diffraction (S-SAD) phasing method. However, the anomalous effect is weak at the wavelengths where data are normally recorded (between 1 and 2 Å) and this limits the potential of this method to well diffracting crystals. Longer wavelengths increase the strength of the anomalous signal but at the cost of increasing air absorption and scatter, which degrade the precision of the anomalous measurement, consequently hindering phase determination. A new instrument, the long-wavelength beamline I23 at Diamond Light Source, was designed to work at significantly longer wavelengths compared with standard synchrotron beamlines in order to open up the native S-SAD method to projects of increasing complexity. Here, the first novel structure, that of the oxidase domain involved in the production of the natural product patellamide, solved on this beamline is reported using data collected to a resolution of 3.15 Å at a wavelength of 3.1 Å. The oxidase is an example of a protein that does not crystallize as the selenium variant and for which no suitable homology model for molecular replacement was available. Initial attempts collecting anomalous diffraction data for native sulfur phasing on a standard macromolecular crystallography beamline using a wavelength of 1.77 Å did not yield a structure. The new beamline thus has the potential to facilitate structure determination by native S-SAD phasing for what would previously have been regarded as very challenging cases with modestly diffracting crystals and low sulfur content.

Publication status:PublishedPeer Review status:Peer reviewedVersion:Publisher's versionDate of acceptance:2016-10-07 Funder: Biotechnology and Biological Research Council   Funder: Royal Society   Funder: European Research Council   Funder: Wellcome Trust   Funder: Sichuan University   Notes:This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.

Bibliographic Details

Publisher: International Union of Crystallography

Publisher Website: http://www.iucr.org/

Journal: Acta Crystallographica Section D: Structural Biologysee more from them

Publication Website: http://journals.iucr.org/d/

Volume: 72

Issue: Pt 11

Issue Date: 2017-10

pages:1174-1180Identifiers

Doi: https://doi.org/10.1107/S2059798316015850

Issn: 2059-7983

Issn: 2059-7983

Uuid: uuid:36986752-fbc0-4338-8505-9c9fcf0c662a

Urn: uri:36986752-fbc0-4338-8505-9c9fcf0c662a

Pubs-id: pubs:699009 Item Description

Type: journal-article;

Language: eng

Version: Publisher's versionKeywords: RIPPs S-SAD azoline oxidase cyanobactins long wavelength phasing structure sulfur

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Author: Bent, AF - - - Mann, G - - - Houssen, WE - - - Mykhaylyk, V - - - Duman, R - - - Thomas, L - - - Jaspars, M - - - Wagner, A - - -

Source: https://ora.ox.ac.uk/objects/uuid:36986752-fbc0-4338-8505-9c9fcf0c662a



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