[1]刘杰,曹志伟,陆晨曦,等.组织蛋白酶C特异性荧光底物的合成和血清学分析应用[J].常州大学学报(自然科学版),2021,33(01):41-47.[doi:10.3969/j.issn.2095-0411.2021.01.007]
　LIU Jie,CAO Zhiwei,LU Chenxi,et al.Synthesis and Serological Analysis of Cathepsin C Specific Fluorescent Substrate[J].Journal of Changzhou University(Natural Science Edition),2021,33(01):41-47.[doi:10.3969/j.issn.2095-0411.2021.01.007]
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组织蛋白酶C特异性荧光底物的合成和血清学分析应用()
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常州大学学报(自然科学版)[ISSN:2095-0411/CN:32-1822/N]

卷:
第33卷
期数:
2021年01期
页码:
41-47
栏目:
生物医药工程
出版日期:
2021-01-20

文章信息/Info

Title:
Synthesis and Serological Analysis of Cathepsin C Specific Fluorescent Substrate
文章编号:
2095-0411(2021)01-0041-07
作者:
刘杰; 曹志伟; 陆晨曦; 戈兴; 周晓鹰
(常州大学 药学院, 江苏 常州 213164)
Author(s):
LIU Jie;  CAO Zhiwei;  LU Chenxi;  GE Xing;  ZHOU Xiaoying
(School of Pharmacy,Changzhou University,Changzhou 213164,China)
关键词:
组织蛋白酶C;  血清检测;  荧光探针
Keywords:
cathepsin C;  serum test;  fluorescent probe
分类号:
R 593.22
DOI:
10.3969/j.issn.2095-0411.2021.01.007
文献标志码:
A
摘要:
组织蛋白酶C是溶酶体半胱氨酸蛋白酶, 源于免疫颗粒细胞。组织蛋白酶C作为近年来的研究热点, 在于其为颗粒细胞炎症性丝氨酸蛋白酶的上游活化酶, 在生理和病理过程中有着重要作用。通过研究设计并合成基于香豆素的荧光底物(CatC-AFC)来评估血清组织蛋白酶C的活性。结果表明, 在特异性底物CatC-AFC与组织蛋白酶C反应后, 荧光强度在非常短的时间内显著增加。酶动力学研究显示组织蛋白酶C以稳健的, 线性的和时间依赖性的方式水解CatC-AFC。通过梯度稀释方法验证基于血清中组织蛋白酶C的活性测定, 并且没有观察到干扰或自发荧光。基于血清中组织蛋白酶C活性的方差系数表明了良好的重现性。
Abstract:
Cathepsin C is a lysosomal cysteine protease derived from immune granulosa cells. As a research hotspot in recent years, cathepsin C is an upstream activating enzyme of granulocyte inflammatory serine protease, which plays an important role in physiological and pathological processes. The coumarin-based fluorogenic substrate(CatC-AFC)was designed and synthesized to evaluate serum cathepsin C activity. The results showed that the fluorescence intensity increased significantly in a very short period of time after the specific substrate CatC-AFC reacted with cathepsin C. Enzymatic kinetic studies showed that cathepsin C hydrolyzed CatC-AFC in a robust, linear, and time-dependent manner. The activity assay based on cathepsin C in serum was verified by a gradient dilution method, and no interference or autofluorescence was observed. The coefficient of variance based on the activity of cathepsin C in serum indicates good reproducibility.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2020-10-07。
基金项目:2019年江苏省研究生科研与实践创新计划项目(KYCX19_1777); 常州大学高层次人才引进启动基金(ZMF14020066); 常州大学科技创新“卓越”项目(Z13)。
作者简介:刘杰(1996—),男,江苏扬州人,硕士生。通信联系人:周晓鹰(1957—),E-mail:xiaoyingzhou@cczu.edu.cn
更新日期/Last Update: 2021-01-20