Proteolytic fingerprinting of complex biological samples using combinatorial libraries of fluorogenic probes.
Proteolytic fingerprinting of complex biological samples using combinatorial libraries of fluorogenic probes.
复制标题
使用荧光探针组合文库对复杂生物样品进行蛋白水解指纹分析。
DOI：
10.1002/0471140864.ps2122s70
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发表时间：
2012
影响因子：
--
通讯作者：
Galande,AmitK
中科院分区：
文献类型：
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作者：
Jambunathan,Kalyani;Watson,DouglasS;Kodukula,Krishna;Galande,AmitK
蛋白酶作为候选生物标志物和许多人类疾病的治疗靶点已经引起了人们的兴趣。一个关键的挑战是在复杂的生物液体环境中识别和表征疾病相关的蛋白酶，例如血清，血浆和支气管肺泡灌洗液，其中多种水解酶协同作用。本单元描述了一个协议，以映射全球蛋白水解底物的复杂的生物样品的特异性，使用一个简洁的组合库的内部淬灭荧光肽探针（IQFP）。这种底物分析方法提供了不同生物样品之间蛋白酶特异性的全局和定量比较。这样的比较分析可以导致识别疾病特异性的蛋白水解活性的“指纹”，其在诊断和治疗中具有潜在的实用性。方案Protein Sci. 70：21.22.1 - 21.22.14。© 2012年由John Wiley & Sons，Inc.
Proteases have garnered interest as candidate biomarkers and therapeutic targets for many human diseases. A key challenge is the identification and characterization of disease‐relevant proteases in the complex milieu of biological fluids such as serum, plasma, and bronchoalveolar lavage, in which a multitude of hydrolases act in concert. This unit describes a protocol to map the global proteolytic substrate specificities of complex biological samples using a concise combinatorial library of internally quenched fluorogenic peptide probes (IQFPs). This substrate profiling approach provides a global and quantitative comparison of protease specificities between different biological samples. Such a comparative analysis can lead to the identification of disease‐specific ‘fingerprints' of proteolytic activities with potential utility in diagnosis and therapy.Curr. Protoc. Protein Sci. 70:21.22.1‐21.22.14. © 2012 by John Wiley & Sons, Inc.