Saidjalolov Saidbakhrom, Chen Xiao-Xiao, Moreno Julia, Cognet Michael, Wong-Dilworth Luis, Bottanelli Francesca, Sakai Naomi, Matile Stefan
Department of Organic Chemistry, University of Geneva, CH-1211 Geneva, Switzerland.
Institute for Chemistry and Biochemistry, Freie Universität Berlin, Thielallee 63, D-14195 Berlin, Germany.
JACS Au. 2024 Aug 20;4(10):3759-3765. doi: 10.1021/jacsau.4c00487. eCollection 2024 Oct 28.
Thiol-mediated uptake (TMU) is thought to occur through dynamic covalent cascade exchange networks. Here we show that the cascade accounting for TMU of asparagusic acid derivatives (AspA) ends in the Golgi apparatus (G) and shifts from disulfide to thioester exchange with palmitoyl transferases as the final exchange partner. As a result, AspA combined with pH-sensitive fluoresceins, red-shifted silicon-rhodamines, or mechanosensitive flipper probes selectively labels the Golgi apparatus in fluorescence microscopy images in living and fixed cells. AspA Golgi trackers work without cellular engineering and excel with speed, simplicity, generality, and compatibility with G/ER and cis/trans discrimination, morphological changes, anterograde vesicular trafficking, and superresolution imaging by stimulated emission depletion microscopy. Golgi flippers in particular can image membrane order and tension in the Golgi and, if desired, at the plasma membrane during TMU.
硫醇介导的摄取（TMU）被认为是通过动态共价级联交换网络发生的。在这里，我们表明，负责天冬氨酸衍生物（AspA）TMU的级联反应在高尔基体（G）中结束，并从二硫键交换转变为与棕榈酰转移酶进行硫酯交换，作为最终的交换伙伴。因此，与pH敏感荧光素、红移硅罗丹明或机械敏感翻转探针结合的AspA在活细胞和固定细胞的荧光显微镜图像中选择性地标记高尔基体。AspA高尔基体追踪器无需细胞工程即可工作，并且在速度、简便性、通用性以及与G/ER和顺/反辨别、形态变化、顺行囊泡运输以及受激发射损耗显微镜超分辨率成像的兼容性方面表现出色。特别是高尔基体翻转探针可以在TMU期间对高尔基体中的膜秩序和张力进行成像，如果需要，还可以对质膜进行成像。
