An approach to quality management in structural biology: Biophysical selection of proteins for successful crystallization

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Item Type:	Article
Title:	An approach to quality management in structural biology: Biophysical selection of proteins for successful crystallization
Creators Name:	Niesen, F.H., Koch, A., Lenski, U., Harttig, U., Roske, Y., Heinemann, U. and Hofmann, K.P.
Abstract:	Aggregation, incorrect folding and low stability are common obstacles for protein structure determination, and are often discovered at a very late state of protein production. In many cases, however, the reasons for failure to obtain diffracting crystals remain entirely unknown. We report on the contribution of systematic biophysical characterization to the success in structural determination of human proteins of unknown fold. Routine analysis using dynamic light scattering (DLS), differential scanning calorimetry (DSC) and Fourier-transform infrared spectroscopy (FTIR) was employed to evaluate fold and stability of 263 purified protein samples (98 different human proteins). We found that FTIR-monitored temperature scanning may be used to detect incorrect folding and discovered a positive correlation between unfolding enthalpy measured with DSC and the size of small, globular proteins that may be used to estimate the quality of protein preparations. Furthermore, our work establishes that the risk of aggregation during concentration of proteins may be reduced through DLS monitoring. In summary, our study demonstrates that biophysical characterization provides an ideal tool to facilitate quality management for structural biology and many other areas of biological research.
Keywords:	Protein Crystallization, Protein Fold, Structural Genomics, Dynamic Light Scattering (DLS), Differential Scanning Calorimetry (DSC), Fourier-Transform Infrared Spectroscopy (FTIR)
Source:	Journal of Structural Biology
ISSN:	1047-8477
Publisher:	Elsevier
Volume:	162
Number:	3
Page Range:	451-459
Date:	June 2008
Official Publication:	https://doi.org/10.1016/j.jsb.2008.03.007
PubMed:	View item in PubMed

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