Robot for lipidic cubic phase (LCP) crystallization of membrane proteins

我们在这里请求一个用于膜蛋白的无规立方相（LCP）结晶的机器人，该机器人将成为蒙特利尔大学生物化学系结构生物学（FSB）机构设施的一部分。由于它们的疏水性，很难过量生产和纯化膜蛋白用于结构分析。在过去5-10年中，使用不同的去污剂筛选、稳定化策略（例如与稳定化结构域融合或通过抗抗体或纳米抗体）的进展已经使得一些膜蛋白可用于X射线晶体学分析。一种特别成功的方法是在LCP或内消旋中结晶，其中膜蛋白在与膜相似的凝胶状脂质基质中结晶。在LCP中生长的蛋白质晶体通过亲水和疏水部分进行接触，从而导致较低的溶剂含量和更好的晶体。然而，蒙特利尔大学、麦吉尔大学和康考迪亚大学现有的机器人设备不允许筛选LCP结晶条件，因为现有的机器人无法处理LCP溶液的极端粘度（通常与牙膏相比）。这限制了正在进行的膜蛋白结晶研究，不仅在我们的机构，但在蒙特利尔的整个结构生物学研究社区。从Concordia和麦吉尔的主要结构生物学家是共同申请人这一事实反映了收购这种仪器的必要性和紧迫性。LCP机器人将很容易进入蒙特利尔的整个结构生物学研究社区。

We are here requesting a robot for lipidic cubic phase (LCP) crystallization of membrane proteins that will be part of the CFI-funded institutional Facility for Structural Biology (FSB) in the Biochemistry Department at the Université de Montréal. Due to their hydrophobic nature, it is extremely difficult to overproduce and purify membrane proteins for structural analysis. Progress over the last 5-10 years using different detergent screens, stabilization strategies (e.g. fusion with stabilizing domains or by anti- or nanobodies) have rendered some membrane proteins accessible to X-ray crystallographic analysis. A particularly successful approach is the crystallization in LCP or in meso in which membrane proteins are crystallized in a gel-like lipid-based matrix that bears similarity to the membrane. Protein crystals grown in LCP make contacts through hydrophilic as well as hydrophobic portions resulting in lower solvent content and better crystals. However, the existing robotic equipment at Université de Montréal, McGill and Concordia does not allow the screening of LCP crystallization conditions since the available robots can not handle the extreme viscosity of the LCP solutions (often compared to tooth paste). This limits the ongoing research on crystallization of membrane proteins, not only in our institution, but in the entire structure biology research community in Montréal. The need and urgency of the acquisition of this instrumentation are reflected by the fact that leading structural biologists from Concordia and McGill are co-applicants. The LCP robot will be readily accessible to the entire structure biology research community in Montréal.