TY - JOUR
T1 - Macro-to-micro structural proteomics
T2 - Native source proteins for high-throughput crystallization
AU - Totir, Monica
AU - Echols, Nathaniel
AU - Nanao, Max
AU - Gee, Christine L.
AU - Moskaleva, Alisa
AU - Gradia, Scott
AU - Iavarone, Anthony T.
AU - Berger, James M.
AU - May, Andrew P.
AU - Zubieta, Chloe
AU - Alber, Tom
PY - 2012/2/29
Y1 - 2012/2/29
N2 - Structural biology and structural genomics projects routinely rely on recombinantly expressed proteins, but many proteins and complexes are difficult to obtain by this approach. We investigated native source proteins for high-throughput protein crystallography applications. The Escherichia coli proteome was fractionated, purified, crystallized, and structurally characterized. Macro-scale fermentation and fractionation were used to subdivide the soluble proteome into 408 unique fractions of which 295 fractions yielded crystals in microfluidic crystallization chips. Of the 295 crystals, 152 were selected for optimization, diffraction screening, and data collection. Twenty-three structures were determined, four of which were novel. This study demonstrates the utility of native source proteins for high-throughput crystallography.
AB - Structural biology and structural genomics projects routinely rely on recombinantly expressed proteins, but many proteins and complexes are difficult to obtain by this approach. We investigated native source proteins for high-throughput protein crystallography applications. The Escherichia coli proteome was fractionated, purified, crystallized, and structurally characterized. Macro-scale fermentation and fractionation were used to subdivide the soluble proteome into 408 unique fractions of which 295 fractions yielded crystals in microfluidic crystallization chips. Of the 295 crystals, 152 were selected for optimization, diffraction screening, and data collection. Twenty-three structures were determined, four of which were novel. This study demonstrates the utility of native source proteins for high-throughput crystallography.
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U2 - 10.1371/journal.pone.0032498
DO - 10.1371/journal.pone.0032498
M3 - Article
C2 - 22393408
AN - SCOPUS:84857664097
SN - 1932-6203
VL - 7
JO - PloS one
JF - PloS one
IS - 2
M1 - e32498
ER -