Improving macromolecular crystallization using lattice-promoting variants of GFP The bottleneck in structure determination by X-ray crystallography is crystallization, where -70% of purified proteins fail. A major reason proteins fail to crystallize is insufficient suitable lattice-forming contacts. The central hypothesis in this project is that the probability of obtaining crystals is increased every time a new form of a molecule with new potential crystal contacts is tested. We will develop rapid and general methods to generate numerous venations on a protein molecule by linking the protein to a series of oligomeric forms of green fluorescent protein (GFP). The use of GFP is a major advantage because it can be linked firmly to practically any protein by inserting a hairpin (two strands connected by a loop) from GFP into a loop of the target protein. The target protein containing this extra hairpin can bind tightly to a

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
5P01GM098177-02
Application #
8471726
Study Section
Special Emphasis Panel (ZRG1-BCMB-A)
Project Start
Project End
Budget Start
2013-06-01
Budget End
2014-05-31
Support Year
2
Fiscal Year
2013
Total Cost
$509,364
Indirect Cost
$203,623
Name
Los Alamos National Lab
Department
Type
DUNS #
175252894
City
Los Alamos
State
NM
Country
United States
Zip Code
87545
Close, Devin W; Paul, Craig Don; Langan, Patricia S et al. (2014) Thermal green protein, an extremely stable, nonaggregating fluorescent protein created by structure-guided surface engineering. Proteins :
Hart, Darren J; Waldo, Geoffrey S (2013) Library methods for structural biology of challenging proteins and their complexes. Curr Opin Struct Biol 23:403-8