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dc.contributor.authorShaw, Neil
dc.contributor.authorCheng, Chongyun
dc.contributor.authorTempel, Wolfram
dc.contributor.authorChang, Jessie
dc.contributor.authorNg, Joseph
dc.contributor.authorWang, Xin-Yu
dc.contributor.authorPerrett, Sarah
dc.contributor.authorRose, John
dc.contributor.authorRao, Zihe
dc.contributor.authorWang, Bi-Cheng
dc.contributor.authorLiu, Zhi-Jie
dc.date.accessioned2013-06-12T15:16:13Z
dc.date.available2013-06-12T15:16:13Z
dc.date.issued2007-07-07
dc.identifier.citationBMC Structural Biology. 2007 Jul 07;7(1):46
dc.identifier.urihttp://dx.doi.org/10.1186/1472-6807-7-46
dc.identifier.urihttp://hdl.handle.net/10724/19754
dc.description.abstractAbstract Background The major bottleneck for determination of 3 D structures of proteins using X-rays is the production of diffraction quality crystals. Often proteins are subjected to chemical modification to improve the chances of crystallization Results Here, we report the successful crystallization of a nuclease employing a reductive methylation protocol. The key to crystallization was the successful introduction of 44 new cohesive (NZ) CH...O contacts (3.2 – 3.7 Å) by the addition of 2 methyl groups to the side chain amine nitrogen (NZ) of 9 lysine residues of the nuclease. The new contacts dramatically altered the crystallization properties of the protein, resulting in crystals that diffracted to 1.2 Å resolution. Analytical ultracentrifugation analysis and thermodynamics results revealed a more compact protein structure with better solvent exclusion of buried Trp residues in the folded state of the methylated protein, assisting crystallization. Conclusion In this study, introduction of novel cohesive (NZ)CH...O contacts by reductive methylation resulted in the crystallization of a protein that had previously resisted crystallization in spite of extensive purification and crystallization space screening. Introduction of (NZ)CH...O contacts could provide a solution to crystallization problems for a broad range of protein targets.
dc.title(NZ)CH...O Contacts Assist Crystallization of a ParB-like Nuclease
dc.typeJournal Article
dc.date.updated2013-06-07T19:11:41Z
dc.description.versionPeer Reviewed
dc.language.rfc3066en
dc.rights.holderNeil Shaw et al.; licensee BioMed Central Ltd.


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