Activatable endoplasmic reticulum-targeted NIR fluorescent probe with a large Stokes shift for detecting and imaging chymotrypsin,Analyst

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Activatable endoplasmic reticulum-targeted NIR fluorescent probe with a large Stokes shift for detecting and imaging chymotrypsin
Analyst ( IF 3.6 ) Pub Date : 2022-08-08 , DOI: 10.1039/d2an01013e
Ting Lan ₁ , Qin-Qin Tian ₁ , Ming-Hua Li ₁ , Wei He ₁

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In this work, the first endoplasmic reticulum-targeted near-infrared fluorescent probe, ISO-Chy, with a dicyanoisophorone derivative as a fluorophore is reported by introducing the recognition group of 4-bromobutyl for chymotrypsin detection. The probe can be easily synthesized and has shown satisfactory sensitivity and selectivity to chymotrypsin. Meanwhile, ISO-Chy has a large Stokes shift (135 nm) to minimize self-absorption and interference from autofluorescence and then generate significant fluorescence enhancement upon incubation with chymotrypsin. Additionally, ISO-Chy has an excellent ability to target the endoplasmic reticulum, along with preferable Pearson's correlation coefficients (R_r) of 0.9411 and 0.9522 in P815 cells and HepG2 cells, respectively. Moreover, ISO-Chy was successfully utilized to visualize endogenous chymotrypsin in P815 cells and HepG2 cells and was first used to detect chymotrypsin activity in HepG2 tumor-bearing mice. These findings indicate that ISO-Chy could be an effective tool for detecting endogenous chymotrypsin activity, supporting its use for investigating chymotrypsin function in pathologic processes.

中文翻译：

具有大斯托克斯位移的可激活内质网靶向 NIR 荧光探针，用于检测和成像糜蛋白酶

在这项工作中，通过引入用于胰凝乳蛋白酶检测的 4-溴丁基识别基团，报道了第一个以二氰基异佛尔酮衍生物为荧光团的内质网靶向近红外荧光探针 ISO-Chy。该探针易于合成，对糜蛋白酶表现出令人满意的灵敏度和选择性。同时，ISO-Chy 具有较大的斯托克斯位移 (135 nm)，可最大限度地减少自体荧光的自吸收和干扰，然后在与糜蛋白酶孵育时产生显着的荧光增强。此外，ISO-Chy 具有出色的靶向内质网的能力，同时具有较好的 Pearson 相关系数 ( R _r) 在 P815 细胞和 HepG2 细胞中分别为 0.9411 和 0.9522。此外，ISO-Chy 成功地用于观察 P815 细胞和 HepG2 细胞中的内源性糜蛋白酶，并首次用于检测 HepG2 荷瘤小鼠的糜蛋白酶活性。这些发现表明 ISO-Chy 可能是检测内源性胰凝乳蛋白酶活性的有效工具，支持其用于研究病理过程中的胰凝乳蛋白酶功能。

更新日期：2022-08-10

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