Abstract:
Conditional depletion of proteins is a potential strategy to elucidate protein function, especially in complex cellular processes like meiosis. Several methods are available to effectively deplete a protein in a conditional manner. Conditional loss of a protein function can be achieved by depleting it from its region of action by degrading it. A conditional loss of protein function can also be achieved by sequestering it to a functionally unavailable compartment inside the cell. This chapter describes anchor away, a conditional depletion tool that can deplete a protein both temporally and spatially by translocation. It utilizes the affinity of FRB to bind FKBP12 in the presence of rapamycin for a quick and efficient translocation of the protein to a designated location. Anchor away is a reliable tool for the study of meiotic proteins, as only small quantities of rapamycin are required to efficiently and rapidly translocate the protein of interest without compromising meiotic progression.
摘要:
蛋白质的条件消耗是阐明蛋白质功能的潜在策略,尤其是在复杂的细胞过程中,如减数分裂。有几种方法可用于以条件方式有效地耗尽蛋白质。蛋白质功能的有条件丧失可以通过降解使其从其作用区域耗尽来实现。蛋白质功能的条件性丧失也可以通过将其隔离到细胞内功能不可用的区室来实现。本章描述了锚远,一种有条件的消耗工具,可以通过易位在时间和空间上消耗蛋白质。它利用FRB的亲和力在雷帕霉素的存在下结合FKBP12,将蛋白质快速有效地转移到指定位置。锚定去除是减数分裂蛋白研究的可靠工具,因为只需要少量的雷帕霉素就可以有效和快速地转移目的蛋白而不影响减数分裂进程。