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

Monica Totir, Nathaniel Echols, Max Nanao, Christine L. Gee, Alisa Moskaleva, Scott Gradia, Anthony T. Iavarone, James M. Berger, Andrew P. May, Chloe Zubieta, Tom Alber

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Article numbere32498
JournalPloS one
Volume7
Issue number2
DOIs
StatePublished - Feb 29 2012

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Fingerprint Dive into the research topics of 'Macro-to-micro structural proteomics: Native source proteins for high-throughput crystallization'. Together they form a unique fingerprint.

  • Cite this

    Totir, M., Echols, N., Nanao, M., Gee, C. L., Moskaleva, A., Gradia, S., Iavarone, A. T., Berger, J. M., May, A. P., Zubieta, C., & Alber, T. (2012). Macro-to-micro structural proteomics: Native source proteins for high-throughput crystallization. PloS one, 7(2), [e32498]. https://doi.org/10.1371/journal.pone.0032498