Abstract
The inclusion of protein contaminants into crystals of turkey egg white lysozyme (TEWL) was investigated by electrospray mass spectrometry of the dissolved crystals. The results show that significant amounts of the structurally related contaminant hen egg white lysozyme (HEWL) are included in the crystals of TEWL. The structurally unrelated contaminant RNAse A, on the other hand, is not included. The X-ray diffraction data statistics of a hybrid TEWL/HEWL crystal and an uncontaminated TEWL crystal were of similar quality. This indicates that, even though the crystals contain much higher levels of the contaminant than one would have expected after a recrystallization experiment, they are still suitable for X-ray diffraction experiments. However, attempts to detect the presence of the contaminant in the crystal by crystallographic structure refinement did not yield conclusive results.
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Selected References
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- Bott R., Sarma R. Crystal structure of turkey egg-white lysozyme: results of the molecular replacement method at 5 A resolution. J Mol Biol. 1976 Oct 5;106(4):1037–1046. doi: 10.1016/0022-2836(76)90351-x. [DOI] [PubMed] [Google Scholar]
- Fontecilla-Camps J. C., de Llorens R., le Du M. H., Cuchillo C. M. Crystal structure of ribonuclease A.d(ApTpApApG) complex. Direct evidence for extended substrate recognition. J Biol Chem. 1994 Aug 26;269(34):21526–21531. doi: 10.2210/pdb1rcn/pdb. [DOI] [PubMed] [Google Scholar]
- Hirschler J., Fontecilla-Camps J. C. Contaminant effects on protein crystal morphology in different growth environments. Acta Crystallogr D Biol Crystallogr. 1996 Jul 1;52(Pt 4):806–812. doi: 10.1107/S0907444996001813. [DOI] [PubMed] [Google Scholar]
- Siuzdak G. The emergence of mass spectrometry in biochemical research. Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11290–11297. doi: 10.1073/pnas.91.24.11290. [DOI] [PMC free article] [PubMed] [Google Scholar]