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Calorimetric approaches to characterizing effects of additives on protein crystallization

机译：量热法表征添加剂对蛋白质结晶的影响

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Calorimetric Approaches to Introduction of additives to the crystallization solution can improve the quality of macromolecular crystals, presumably by enhancing crystal contacts, conformational stability, and/or solvent interactions. However, despite their widespread use, the mechanisms by which these additives exert their effects remain largely unknown. We have used differential scanning calorimetry and isothermal titration calorimetry (ITC) to characterize the effects of additives on the crystallization of several proteins. We have found that the positive effects on crystallizability exerted by an additive can be correlated with stabilization of the macromolecule in solution, as evidenced by an increase in the midpoint melting transition temperature. Additionally, the additives' effect on the thermodynamic variables Delta H and Delta G, as determined by ITC, can be used to distinguish whether the additive functions by binding to the macromolecule or by altering the properties of the crystallization solution. The use of ultrasensitive instrumental methods to evaluate the potential effectiveness of additives can facilitate, rapid screening of a large number of additives prior to crystallization and provide a means to rationally limit the crystallization solution search space.
机译：将添加剂引入结晶溶液的量热方法可以改善大分子晶体的质量，大概是通过增强晶体接触，构象稳定性和/或溶剂相互作用。然而，尽管它们被广泛使用，但是这些添加剂发挥其作用的机理仍然很大程度上未知。我们已经使用差示扫描量热法和等温滴定量热法（ITC）来表征添加剂对几种蛋白质结晶的影响。我们已经发现，添加剂对结晶性的积极影响可以与溶液中大分子的稳定相关，如中点熔融转变温度的升高所证明的。另外，由ITC确定的添加剂对热力学变量Delta H和Delta G的影响可用于区分添加剂是通过结合大分子还是通过改变结晶溶液的性质起作用。使用超灵敏的仪器方法来评估添加剂的潜在效力可以促进在结晶之前快速筛选大量添加剂，并提供一种合理限制结晶溶液搜索空间的方法。

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