CRYSTALLIZATION IN LOOPS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 蛋白质晶体的可用性是从X射线衍射实验中确定蛋白质三维结构的先决条件。蛋白质晶体通常生长在悬挂式或坐式液滴中，通常被转移到环形或微型支架上进行冷冻和数据收集。从水滴中转移晶体是一件很棘手的事情。通常会影响晶体质量，有时还会导致晶体丢失的过程。然而，从水滴中去除晶体的方法还没有被自动化，因此已经提出了一些替代方案来绕过这个问题。我们已经开发出一种在冷冻环或微支架中生长蛋白质晶体的方法。直接在冷冻环或微支架中生长晶体有几个优点：a)它有助于晶体处理的自动化；b)在某些情况下，它提高了衍射能力的质量(正如我们能够从BL9-1和BL9-2上测量的数据所证明的那样)；以及c)可以在非常小的溶液体积中获得更高水平的过饱和度，从而能够探索额外的结晶条件。该方法还将允许对通过其他方法生长的晶体进行有效的脱水。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The availability of protein crystals is a prerequisite to the determination of the three dimensional structure of proteins from x-ray diffraction experiments. Protein crystals are usually grown in hanging or sitting drops and generally get transferred to a loop or micromount for cryo-freezing and data collection. The transfer of crystals from drops is a ?tricky? process that often affects the quality of crystals and sometimes results in the loss of the crystal. However, the method of removing a crystal from a drop has not been automated, so a number of alternatives have been suggested to circumvent this problem. We have developed a method to grow protein crystals in cryoloops or micromounts. Growing crystals directly in cryoloops or micromounts has several advantages: a) it facilitates the automation of crystal handling; b) in some cases, it improves the quality of the diffraction ability (as we have been able to demonstrate from data measured on BL9-1 and BL9-2); and c) higher levels of supersaturation can be obtained in very small solution volumes enabling the exploration of additional crystallization conditions. The method will also allow efficient dehydration of crystals grown by other methods.