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Cascade-Activated AIEgen-Peptide Probe for Noninvasively Monitoring Chymotrypsin-like Activity of Proteasomes in Cancer Cells
Analytical Chemistry ( IF 6.7 ) Pub Date : 2023-05-30 , DOI: 10.1021/acs.analchem.3c01589
Qishu Jiao 1 , Yaxin Zheng 1 , Shicheng Pei 1 , Xuan Luo 1 , Xiaoxing Wu 2 , Keming Xu 1, 3 , Wenying Zhong 1, 3, 4
Affiliation  

Noninvasive monitoring of chymotrypsin-like (ChT-L) activity of proteasomes is of great significance for the diagnosis and prognosis of various cancers. However, commercially available proteasome probes usually lack adequate cancer-cell selectivity. To noninvasively monitor ChT-L activity of proteasomes in living cells, we rationally designed a cascade-activated AIEgen-peptide probe (abbreviated as TPE-1p), which self-assembled in aqueous solution to exhibit bright fluorescence in response to sequential treatment of alkaline phosphatase (ALP) and ChT-L. Transmission electron microscopy, enzymatic kinetics, and in vitro fluorescence experiments validated that TPE-1p was efficiently dephosphorylated by ALP to generate TPE-1, which was recognized by ChT-L in the proteasome, and transformed to form nanofibers with strong fluorescence signals. Cell imaging experiments revealed that bright blue fluorescence was observed in TPE-1p-treated HeLa cells, whereas NIH3T3 and HepG2 cells showed less fluorescence at the same condition. The enhanced fluorescence signals in HeLa cells were attributed to the high activities of endogenous ALP and ChT-L. Moreover, TPE-1p was utilized to noninvasively assess the inhibition efficiency of a ChT-L inhibitor (bortezomib, abbreviated as Btz) in HeLa cells. Significant correlation was found between the fluorescence signals of TPE and the viabilities of Btz-treated cells in concentration ranges from 0 to 1 μM, indicating that TPE-1p could be employed to predict the activity of ChT-L inhibitors. The design of the cascade-activated AIEgen-peptide probe provides a viable approach for noninvasively monitoring the ChT-L activity of proteasomes in living cells, which facilitates high-throughput screening of ChT-L inhibitors in cancer therapy.

中文翻译:

级联激活的 AIEgen 肽探针用于无创监测癌细胞中蛋白酶体的胰凝乳蛋白酶样活性

无创监测蛋白酶体的胰凝乳蛋白酶样(ChT-L)活性对于各种癌症的诊断和预后具有重要意义。然而,市售的蛋白酶体探针通常缺乏足够的癌细胞选择性。为了无创监测活细胞中蛋白酶体的 ChT-L 活性,我们合理设计了一种级联激活的 AIEgen 肽探针(缩写为 TPE-1p),该探针在水溶液中自组装以响应碱的连续处理而表现出明亮的荧光磷酸酶 (ALP) 和 ChT-L。透射电子显微镜、酶动力学和体外荧光实验验证了 TPE-1p 被 ALP 有效去磷酸化生成 TPE-1,TPE-1 在蛋白酶体中被 ChT-L 识别,并转化形成具有强荧光信号的纳米纤维。细胞成像实验表明,在 TPE-1p 处理的 HeLa 细胞中观察到亮蓝色荧光,而 NIH3T3 和 HepG2 细胞在相同条件下显示出较少的荧光。HeLa 细胞中增强的荧光信号归因于内源性 ALP 和 ChT-L 的高活性。此外,TPE-1p 用于无创评估 ChT-L 抑制剂(硼替佐米,缩写为 Btz)对 HeLa 细胞的抑制效率。TPE 的荧光信号与浓度范围为 0 至 1 μM 的 Btz 处理细胞的活力之间存在显着相关性,表明 TPE-1p 可用于预测 ChT-L 抑制剂的活性。级联激活的 AIEgen 肽探针的设计为无创监测活细胞中蛋白酶体的 ChT-L 活性提供了一种可行的方法,
更新日期:2023-05-30
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