Protein production and crystallization at SECSG - An overview

Bi Cheng Wang, Michael W W Adams, Harry Dailey, Larry DeLucas, Ming Luo, John Rose*, Robert Bunzel, Tamara Dailey, Jeff Habel, Peter Horanyi, Francis E. Jenney, Irina Kataeva, Han Seung Lee, Songlin Li, Ting Li, Dawei Lin, Zhi Jie Liu, Chi Hao Luan, Michael Mayer, Lisa NagyM. Gary Newton, Joseph Ng, Farris L. Poole, Ashit Shah, Claudia Shah, Frank J. Sugar, Hao Xu

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    11 Scopus citations

    Abstract

    Using a high degree of automation, the Southeast Collaboratory for Structural Genomics (SECSG) has developed high throughput pipelines for protein production, and crystallization using a two-tiered approach. Primary, or tier-1, protein production focuses on producing proteins for members of large Pfam families that lack a representative structure in the Protein Data Bank. Target genomes are Pyrococcus furiosus and Caenorhabditis elegans. Selected human proteins are also under study. Tier-2 protein production, or target rescue, focuses on those tier-1 proteins, which either fail to crystallize or give poorly diffracting crystals. This two tier approach is more efficient since it allows the primary protein production groups to focus on the production of new targets while the tier-2 efforts focus on providing additional sample for further studies and modified protein for structure determination. Both efforts feed the SECSG high throughput crystallization pipeline, which is capable of screening over 40 proteins per week. Details of the various pipelines in use by the SECSG for protein production and crystallization, as well as some examples of target rescue are described.

    Original languageEnglish (US)
    Pages (from-to)233-243
    Number of pages11
    JournalJournal of Structural and Functional Genomics
    Volume6
    Issue number2-3
    DOIs
    StatePublished - Sep 2005

    Keywords

    • Crystallization
    • Protein production
    • SECSG
    • Southeast Collaboratory
    • Structural genomics
    • Target salvaging

    ASJC Scopus subject areas

    • Structural Biology
    • Biochemistry
    • Genetics

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