The Integrated Center for Structure and Function Innovation (ISFI) will be a PSI-2 Specialized Center developing and applying a set of synergistic technologies organized to overcome recognized bottlenecks in structure determination at the key steps of production of soluble protein and protein crystallization. The development of these technologies will be embedded in an integrated pipeline for protein structure determination and analysis of function, and the technologies will be applied to difficult proteins and protein complexes from large sequence families, emphasizing proteins with high biological significance. The increased success rates made possible by our technologies will be critical for obtaining structures of proteins and protein complexes of high functional value, both for the PSI and for structural biology as a whole. The increased success rates will lead as well to substantial cost reductions in structure determination. Our application of our methodologies as part of this project will enhance the understanding of protein function and protein-protein interactions relevant to human health. The core technologies to be developed in the ISFI are: (1) systematic construction of protein binding partners for chaperone-assisted crystallography, (2) engineering proteins for crystallization by surface mutagenesis and addition of fusion proteins, (3) directed evolution of proteins for solubility and trapping of protein partners required for folding using one member attached to a reporter protein as a bait, (4) identification of protein partners required for proper folding using informatics approaches confirmed by experiments, and (5) systematic combinatorial crystallization screening and optimization. These technologies will be used to address five major bottlenecks in the structural genomics pipeline: (a) soluble expression, (b) small protein complexes, (c) crystallizability, (d) soluble domains of membrane proteins, and (e) eukaryotic protein structure determination. The technologies will be developed through highly integrated collaborations among 6 institutions. Technologies and results will be made available to other Centers and the broader structural biology community through workshops, publications, personnel exchanges, and the web.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Specialized Center--Cooperative Agreements (U54)
Project #
5U54GM074946-06
Application #
7667972
Study Section
Special Emphasis Panel (ZGM1-CBB-3 (SC))
Program Officer
Basavappa, Ravi
Project Start
2005-07-01
Project End
2011-06-30
Budget Start
2009-07-01
Budget End
2011-06-30
Support Year
6
Fiscal Year
2009
Total Cost
$4,229,449
Indirect Cost
Name
Los Alamos National Lab
Department
Type
DUNS #
175252894
City
Los Alamos
State
NM
Country
United States
Zip Code
87545
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Li, Qufei; Wanderling, Sherry; Sompornpisut, Pornthep et al. (2014) Structural basis of lipid-driven conformational transitions in the KvAP voltage-sensing domain. Nat Struct Mol Biol 21:160-6
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Ru, Heng; Zhao, Lixia; Ding, Wei et al. (2012) S-SAD phasing study of death receptor 6 and its solution conformation revealed by SAXS. Acta Crystallogr D Biol Crystallogr 68:521-30
Li, Qufei; Jogini, Vishwanath; Wanderling, Sherry et al. (2012) Expression, purification, and reconstitution of the voltage-sensing domain from Ci-VSP. Biochemistry 51:8132-42

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