• Login
    View Item 
    •   Athenaeum Home
    • BioMed Central Open Access Articles
    • Open Access Articles by UGA Faculty
    • View Item
    •   Athenaeum Home
    • BioMed Central Open Access Articles
    • Open Access Articles by UGA Faculty
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    (NZ)CH...O Contacts Assist Crystallization of a ParB-like Nuclease

    Thumbnail
    View/Open
    1472-6807-7-46.xml (81.24Kb)
    1472-6807-7-46.pdf (2.495Mb)
    1472-6807-7-46-S1.DOC (35.5Kb)
    1472-6807-7-46-S2.DOC (41Kb)
    Date
    2007-07-07
    Author
    Shaw, Neil
    Cheng, Chongyun
    Tempel, Wolfram
    Chang, Jessie
    Ng, Joseph
    Wang, Xin-Yu
    Perrett, Sarah
    Rose, John
    Rao, Zihe
    Wang, Bi-Cheng
    Liu, Zhi-Jie
    Metadata
    Show full item record
    Abstract
    Abstract 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.
    URI
    http://dx.doi.org/10.1186/1472-6807-7-46
    http://hdl.handle.net/10724/19754
    Collections
    • Open Access Articles by UGA Faculty

    About Athenaeum | Contact Us | Send Feedback
     

     

    Browse

    All of AthenaeumCommunities & CollectionsBy Issue DateAuthorsTitlesSubjectsThis CollectionBy Issue DateAuthorsTitlesSubjects

    My Account

    LoginRegister

    About Athenaeum | Contact Us | Send Feedback