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Macro-to-micro structural proteomics: native source proteins for high-throughput crystallization.

ABSTRACT: 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.

SUBMITTER: Totir M

PROVIDER: S-EPMC3290569 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Macro-to-micro structural proteomics: native source proteins for high-throughput crystallization.

Totir Monica M Echols Nathaniel N Nanao Max M Gee Christine L CL Moskaleva Alisa A Gradia Scott S Iavarone Anthony T AT Berger James M JM May Andrew P AP Zubieta Chloe C Alber Tom T

PloS one 20120229 2

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 ...[more]

PMID: 22393408

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