Download MendeleyProline: Mother Nature's cryoprotectant applied to protein crystallography.
Pemberton, T.A., Still, B.R., Christensen, E.M., Singh, H., Srivastava, D., Tanner, J.J.(2012) Acta Crystallogr D Biol Crystallogr 68: 1010-1018
- PubMed: 22868767
- DOI: https://doi.org/10.1107/S0907444912019580
- Primary Citation of Related Structures:
4E3U, 4E3V, 4E3W, 4E3X - PubMed Abstract:
L-Proline is one of Mother Nature's cryoprotectants. Plants and yeast accumulate proline under freeze-induced stress and the use of proline in the cryopreservation of biological samples is well established. Here, it is shown that L-proline is also a useful cryoprotectant for protein crystallography. Proline was used to prepare crystals of lysozyme, xylose isomerase, histidine acid phosphatase and 1-pyrroline-5-carboxylate dehydrogenase for low-temperature data collection. The crystallization solutions in these test cases included the commonly used precipitants ammonium sulfate, sodium chloride and polyethylene glycol and spanned the pH range 4.6-8.5. Thus, proline is compatible with typical protein-crystallization formulations. The proline concentration needed for cryoprotection of these crystals is in the range 2.0-3.0 M. Complete data sets were collected from the proline-protected crystals. Proline performed as well as traditional cryoprotectants based on the diffraction resolution and data-quality statistics. The structures were refined to assess the binding of proline to these proteins. As observed with traditional cryoprotectants such as glycerol and ethylene glycol, the electron-density maps clearly showed the presence of proline molecules bound to the protein. In two cases, histidine acid phosphatase and 1-pyrroline-5-carboxylate dehydrogenase, proline binds in the active site. It is concluded that L-proline is an effective cryoprotectant for protein crystallography.
Organizational Affiliation:
Department of Chemistry, University of Missouri-Columbia, Columbia, MO 65211, USA.
