Kimmel Jessica, Kehrer Jessica, Frischknecht Friedrich, Spielmann Tobias
Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany.
Integrative Parasitology, Center for Infectious Diseases, Heidelberg University Medical School, Heidelberg, Germany.
Mol Microbiol. 2022 Mar;117(3):553-568. doi: 10.1111/mmi.14815. Epub 2021 Oct 22.
In the last 10 years, proximity-dependent biotinylation (PDB) techniques greatly expanded the ability to study protein environments in the living cell that range from specific protein complexes to entire compartments. This is achieved by using enzymes such as BirA* and APEX that are fused to proteins of interest and biotinylate proteins in their proximity. PDB techniques are now also increasingly used in apicomplexan parasites. In this review, we first give an overview of the main PDB approaches and how they compare with other techniques that address similar questions. PDB is particularly valuable to detect weak or transient protein associations under physiological conditions and to study cellular structures that are difficult to purify or have a poorly understood protein composition. We also highlight new developments such as novel smaller or faster-acting enzyme variants and conditional PDB approaches, providing improvements in both temporal and spatial resolution which may offer broader application possibilities useful in apicomplexan research. In the second part, we review work using PDB techniques in apicomplexan parasites and how this expanded our knowledge about these medically important parasites.
在过去十年中,邻近依赖性生物素化(PDB)技术极大地扩展了研究活细胞中蛋白质环境的能力,其范围从特定的蛋白质复合物到整个细胞区室。这是通过使用诸如BirA*和APEX等与感兴趣的蛋白质融合并对其附近蛋白质进行生物素化的酶来实现的。PDB技术现在也越来越多地用于顶复门寄生虫。在本综述中,我们首先概述主要的PDB方法,以及它们与解决类似问题的其他技术相比如何。PDB对于在生理条件下检测弱的或瞬时的蛋白质相互作用,以及研究难以纯化或蛋白质组成了解甚少的细胞结构特别有价值。我们还强调了新的进展,如新型更小或作用更快的酶变体和条件性PDB方法,这些在时间和空间分辨率上都有所改进,可能为顶复门研究提供更广泛的应用可能性。在第二部分中,我们综述了在顶复门寄生虫中使用PDB技术的工作,以及这如何扩展了我们对这些医学上重要的寄生虫的认识。