Ionic strength reducers: an efficient approach to protein purification and crystallization. Application to two Rop variants.
نویسندگان
چکیده
Detailed knowledge of the influence of various parameters on macromolecular solubility is essential for crystallization. The concept of so-called 'ionic strength reducers' provides insight into the changes in solubility induced by organic solvents and hydrophilic polymers in aqueous electrolytic solutions. A simple and efficient procedure is presented which exploits the properties of ionic strength reducers in the purification and crystallization of proteins. Using two designed variants of the Rop protein as model systems, superior crystals have been obtained compared with conventional techniques. This procedure is particularly useful in cases where excessive nucleation leads to the growth of a large number of tiny crystals that are useless for crystallographic analysis.
منابع مشابه
Solubility, crystallization and chromatographic properties of macromolecules strongly depend on substances that reduce the ionic strength of the solution.
The differential solubility of macromolecules governs their crystallization and is thus of great importance to crystallography (Blundell and Johnson, 1976; McPherson, 1982, 1985a,b, 1990; Feher and Kam, 1985; Giegé and Mokol, 1989; Ollis and White, 1990; Giegé and Ducruix, 1992; Giegé et al., 1994). In addition, precipitation is widely applied as a fractionation method for the purification of m...
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ورودعنوان ژورنال:
- Acta crystallographica. Section D, Biological crystallography
دوره 60 Pt 7 شماره
صفحات -
تاریخ انتشار 2004