Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 102, Issue 6, Pages 1901-1905Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0408490102
Keywords
crystallographic resolution; NMR structure; x-ray structure; unfolded proteins
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Funding
- NIGMS NIH HHS [P50 GM062411, GM062411] Funding Source: Medline
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In the Joint Center for Structural Genomics, one-dimensional (1D) H-1 NMR spectroscopy is routinely used to characterize the folded state of protein targets and, thus, serves to guide subsequent crystallization efforts and to identify proteins for NMR structure determination. Here, we describe 1D H-1 NMR screening of a group of 79 mouse homologue proteins, which correlates the NMR data with the outcome of subsequent crystallization experiments and crystallographic structure determination. Based on the 1D H-1 NMR spectra, the proteins are classified into four groups, A to D. A-type proteins are candidates for structure determination by NMR or crystallography; B-type are earmarked for crystallography; C indicates folded globular proteins with broadened line shapes; and D are nonglobular, unfolded polypeptides. The results obtained from coarse- and fine-screen crystallization trials imply that only A- and B-type proteins should be used for extensive crystallization trials in the future, with C and D proteins subjected only to coarse-screen crystallization trials. Of the presently studied 79 soluble protein targets, 63% yielded A- or B-quality 1D H-1 NMR spectra. Although similar yields of crystallization hits were obtained for all four groups, A to D, crystals from A- and B-type proteins diffracted on average to significantly higher resolution than crystals produced from C- or D-type proteins. Furthermore, the output of refined crystal structures from this test set of proteins was 4-fold higher for A- and B-type than for C- and D-type proteins.
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