Protein production and crystallization at SECSG -- an overview.
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.
Wang
Bi-Cheng
BC
Southeast Collaboratory for Structural Genomics, University of Georgia, Athens, GA 30602, USA.
Adams
Michael W W
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Dailey
Harry
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DeLucas
Larry
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Luo
Ming
M
Rose
John
J
Bunzel
Robert
R
Dailey
Tamara
T
Habel
Jeff
J
Horanyi
Peter
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Jenney
Francis E
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Kataeva
Irina
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Lee
Han-Seung
HS
Li
Songlin
S
Li
Ting
T
Lin
Dawei
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Liu
Zhi-Jie
ZJ
Luan
Chi-Hao
CH
Mayer
Michael
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Nagy
Lisa
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Newton
M Gary
MG
Ng
Joseph
J
Poole
Farris L
FL
Shah
Ashit
A
Shah
Claudia
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Sugar
Frank J
FJ
Xu
Hao
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D000818:Animals; D017173:Caenorhabditis elegans / Q000737:chemistry*; D003460:Crystallization / Q000379:methods*; D030562:Databases, Protein; D006801:Humans; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D040901:Proteomics / Q000295:instrumentation / Q000379:methods* / Q000458:organization & administration; D020027:Pyrococcus furiosus / Q000737:chemistry*
D011506:Proteins