Salim Aleksandar, Werther Philipp, Hatzopoulos Georgios N, Reymond Luc, Wombacher Richard, Gönczy Pierre, Johnsson Kai
Department of Chemical Biology, Max Planck Institute for Medical Research, Jahnstrasse 29, Heidelberg 69120, Germany.
Institute of Chemical Sciences and Engineering (ISIC), École Polytechnique Fédérale de Lausanne (EPFL), Lausanne 1015, Switzerland.
JACS Au. 2023 Aug 4;3(8):2247-2256. doi: 10.1021/jacsau.3c00271. eCollection 2023 Aug 28.
Polo-like kinase (Plk4) is a serine/threonine-protein kinase that is essential for biogenesis of the centriole organelle and is enriched at centrioles. Herein, we introduce Cen-TCO, a chemical probe based on the Plk4 inhibitor centrinone, to image Plk4 and centrioles in live or fixed cultured human cells. Specifically, we established a bio-orthogonal two-step labeling system that enables the Cen-TCO-mediated imaging of Plk4 by STED super-resolution microscopy. Such direct labeling of Plk4 results in an increased resolution in STED imaging compared with using anti-Plk4 antibodies, underlining the importance of direct labeling strategies for super-resolution microscopy. We anticipate that Cen-TCO will become an important tool for investigating the biology of Plk4 and of centrioles.
Polo样激酶（Plk4）是一种丝氨酸/苏氨酸蛋白激酶，对中心粒细胞器的生物发生至关重要，且在中心粒处富集。在此，我们引入Cen-TCO，一种基于Plk4抑制剂中心体素的化学探针，用于对活的或固定的培养人细胞中的Plk4和中心粒进行成像。具体而言，我们建立了一种生物正交两步标记系统，该系统能够通过受激发射损耗（STED）超分辨率显微镜对Plk4进行Cen-TCO介导的成像。与使用抗Plk4抗体相比，这种对Plk4的直接标记在STED成像中提高了分辨率，突出了直接标记策略对超分辨率显微镜的重要性。我们预计Cen-TCO将成为研究Plk4和中心粒生物学的重要工具。
