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使用无活性寡肽酶的底物捕获分析来鉴定新型肽

Substrate Capture Assay Using Inactive Oligopeptidases to Identify Novel Peptides.

作者信息

Rioli Vanessa, Ferro Emer S

机构信息

Special Laboratory of Applied Toxinology (LETA), Center of Toxins, Immune Response and Cell Signaling (CETICS), Butantan Institute, São Paulo, SP, 05503-000, Brazil.

Department of Pharmacology, Biomedical Science Institute, University of São Paulo, Av. Prof. Lineu Prestes 1524, Sala 317, São Paulo, SP, 05508-000, Brazil.

出版信息

Methods Mol Biol. 2018;1719:97-105. doi: 10.1007/978-1-4939-7537-2_6.

Abstract

Researchers are always searching for novel biologically active molecules including peptides. With the improvement of equipment for electrospray mass spectrometry, it is now possible to identify hundreds of novel peptides in a single run. However, after identifying the peptide sequences it is expensive to synthesize all the peptides to perform biological activity assays. Here, we describe a substrate capture assay that uses inactive oligopeptidases to identify putative biologically active peptides in complexes peptide mixtures. This methodology can use any crude extracts of biological tissues or cells, with the advantage to introduce a filter (i.e., binding to an inactive oligopeptidase) as a prior step in screening to bioactive peptides.

摘要

研究人员一直在寻找包括肽在内的新型生物活性分子。随着电喷雾质谱设备的改进,现在有可能在一次运行中鉴定出数百种新型肽。然而,在鉴定出肽序列后,合成所有肽以进行生物活性测定成本很高。在这里,我们描述了一种底物捕获测定法,该方法使用无活性的寡肽酶来鉴定复合肽混合物中假定的生物活性肽。这种方法可以使用生物组织或细胞的任何粗提物,其优点是在筛选生物活性肽之前引入一个过滤器(即与无活性的寡肽酶结合)作为步骤。

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