Hirschler J, Charon M H, Fontecilla-Camps J C
Laboratoire de Cristallographie et de Cristallogénèse des Protéines, Institut de Biologie Structurale Jean-Pierre Ebel (CEA-CNRS), Grenoble, France.
Protein Sci. 1995 Dec;4(12):2573-7. doi: 10.1002/pro.5560041214.
Filtration effects of turkey egg white lysozyme solution (TEWL) prior to subjecting it to crystallization conditions are investigated. Filtering TEWL solution and crystallizing it in ungelled media significantly decreased the number of conditions yielding crystals. This decrease dependent on the membrane cut-off used for filtration. From this, the postulated factors aiding in nucleation are estimated to be 0.17 microns in diameter. The existence of these factors was verified by the procedure of reversed filtration: filtered solutions passed through their inverted filter membrane a second time lead to improved crystallization results. The effect of aging of the TEWL solution prior to subjecting it to ungelled crystallization conditions was also verified. We did not find any time-dependent change in the size or the number of crystals per drop. Repeating the filtration experiments in agarose-gelled crystallization media showed that the influence of filtration on the crystallization outcome was significantly diminished. Far better crystallization results were obtained compared to ungelled media. However, there is a certain aging effect linked to filtration in gelled media. Different crystallization results were obtained depending on whether filtration was performed before or after aging and subsequent crystallization. This suggests a secondary time-dependent effect.
研究了火鸡蛋白溶菌酶溶液（TEWL）在结晶条件下的过滤效果。在未凝胶化的介质中过滤TEWL溶液并使其结晶，显著减少了产生晶体的条件数量。这种减少取决于用于过滤的膜截留分子量。据此，推测有助于成核的因素直径估计为0.17微米。通过反向过滤程序验证了这些因素的存在：过滤后的溶液再次通过其倒置的滤膜会导致结晶结果得到改善。还验证了TEWL溶液在未凝胶化结晶条件下老化的影响。我们没有发现每滴晶体的大小或数量随时间的变化。在琼脂糖凝胶化结晶介质中重复过滤实验表明，过滤对结晶结果的影响显著减小。与未凝胶化介质相比，获得了更好的结晶结果。然而，在凝胶化介质中过滤存在一定的老化效应。根据过滤是在老化和随后结晶之前还是之后进行，会得到不同的结晶结果。这表明存在二次时间依赖性效应。
