跳至主要内容

Peptide cleavage induced assembly enables highly sensitive electrochemiluminescence detection of protease activity

文献作者：Meng Cheng, Jun Zhou, Xiaoming Zhou, Da Xing   《Sensors and Actuators B: Chemical》

ABSTRACTS
Proteases perform essential functions in a multitude of physiological processes and participate in many human diseases. Probing protease activity sensitively and accurately is critical for both basic research and clinical diagnosis. Herein, a novel optical biosensor for high-sensitive detection of protease was developed based on the specific protease cleavage of synthesized peptide substrate and subsequent assembly between exposed cysteine and Ru(bpy)32+-2-cyanobenzothiazole (Ru(bpy)32+-CBT). The assembly complexes can be enriched by streptavidin coated magnetic beads on work electrode for high-sensitive electrochemiluminescence (ECL) analysis. Using trypsase and caspase-3 as the examples, the limit of detection (LOD) of 8.4?×?10?4?U?mL?1 and 5?×?10?6?U?mL?1 were obtained respectively. This represents one of most sensitive protease assay reported. Current peptide cleavage induced assembly (PCIA) method can be easily extended to detect other proteases by changing peptide substrate sequence, shows its great potential as versatile biosensor in sensing techniques and clinical diagnosis.
    KEY WORDS
    Electrochemiluminescence
    Ru(bpy)32+-2-cyanobenzothiazole (Ru(bpy)32+-CBT)
    Peptide cleavage induced assembly
    Trypsase
    Caspase-3
    SCREENSHOT

    RELATED PRODUCTS
    Peptides were synthesized by ChinaPeptides Co., Ltd (Shanghai, China).
    CHAINING
    https://www.sciencedirect.com/science/article/pii/S0925400518302156
    标签：

    评论

    发表评论

    此博客中的热门博文

    The Catalytically Inactive Mutation of the Ubiquitin-Conjugating Enzyme CDC34 Affects its Stability and Cell Proliferation

    Author：Xun Liu, Yang Zhang, Zhanhong Hu, Qian Li, Lu Yang,  Guoqiang Xu     《The Protein Journal》 ABSTRACTS The ubiquitin proteasome system (UPS) plays important roles in the regulation of protein stability, localization, and activity. A myriad of studies have focused on the functions of ubiquitin ligases E3s and deubiquitinating enzymes DUBs due to their specificity in the recognition of downstream substrates. However, the roles of the most ubiquitin-conjugating enzymes E2s are not completely understood except that they transport the activated ubiquitin and form E2–E3 protein complexes. Ubiquitin-conjugating enzyme CDC34 can promote the degradation of downstream targets through the UPS whereas its non-catalytic functions are still elusive. Here, we find that mutation of the catalytically active cysteine to serine (C93S) results in the reduced ubiquitination, increased stability, and attenuated degradation rate of CDC34. Through semi-quantitative proteom...
    发表评论
    阅读全文

    Identification and Inhibitory Mechanism of Angiotensin I-Converting Enzyme Inhibitory Peptides Derived from Bovine Hemoglobin

    Author：Ying Wang, Yiqun Jiang, Yongguang Yin, Jiyun Liu,  Long Ding, Jingbo Liu, Ting Zhang     《The Protein Journal》 ABSTRACTS Angiotensin I-converting enzyme (ACE, EC.3.4.15.1) inhibitory peptide is an efficacious therapy for hypertension. In this study, four dipeptides, TY, FD, FL and FG, were identified from the desalted fraction of bovine hemoglobin hydrolysate, obtained by in vitro simulated gastrointestinal digestion, via liquid chromatography-tandem mass spectrometry (LC-MS/MS). The IC 50   value of TY and FL are 96.43?±?6.17 and 290.66?±?57.92  μM, respectively. The result of molecular docking indicated that TY occupied the ACE subsite S1 and S1′ with a lowest estimated binding energy of ?9.96  Kcal/mol, while FL occupied the subsite S5 with a lowest estimated binding energy of ?9.37  Kcal/mol. The subsite S1′ and S2′ are closer to the ACE active center (Zn 2+ ) than S5, and the lowest estimated binding energy of TY is lower ...
    发表评论
    阅读全文

    Preparation of an antimicrobial surface by direct assembly of antimicrobial peptide with its surface binding activity

    Author：Junjian Chen, Yuchen Zhu, Yancheng Song, Lin Wang,  Jiezhao Zhan, Jingcai He, Jian Zheng, Chunting Zhong, Xuetao Shi, Sa Liu, Li Ren and Yingjun Wang     《Journal of Materials Chemistry B》 ABSTRACTS Antimicrobial peptides (AMPs) are a broad prospect for clinical application against bacterial infections of biomaterials. However, a bottleneck exists as there is a lack of simple technology to prepare AMPs on biomaterials with sufficient activity, as the activity of AMP is dependent on the correct orientation on the biomaterial. In the present study, based on the conventional AMP (Tet213: KRWWKWWRRC) and surface binding peptide (SKHKGGKHKGGKHKG), we designed an Anchor-AMP that could be directly assembled onto the surface of the biomaterial and also showed excellent antimicrobial activity. By characterizing the surface using a quartz crystal microbalance with dissipation (QCM-D), contact angle, atom force microscopy (AFM) and X-ray photoelectron spectr...
    发表评论
    阅读全文