Recent advances and concepts in substrate specificity determination of proteases using tailored libraries of fluorogenic substrates with unnatural amino acids

Recent advances and concepts in substrate specificity determination of proteases using tailored libraries of fluorogenic substrates with unnatural amino acids
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使用非天然氨基酸荧光底物定制文库测定蛋白酶底物特异性的最新进展和概念

DOI:
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发表时间:
2015
影响因子:
3.7
通讯作者:
M. Drąg
M. Drąg
中科院分区:
生物学2区
文献类型:
--
作者:
Wioletta Rut;Paulina Kasperkiewicz;A. Byzia;M. Poręba;Katarzyna Groborz;M. Drąg

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摘要蛋白酶底物特异性的测定有多种方法,其中最常用的是位置扫描文库法、蛋白质组学法和噬菌体展示法。经典的方法可以提供关于几乎所有蛋白酶结合口袋中天然氨基酸偏好的信息。然而,最近的研究表明,通过将非天然氨基酸应用于位置扫描文库方法,能够获得更多的信息。这些知识可用于设计更有活性和特异性的底物、抑制剂和基于活性的探针。在这篇小综述中,我们描述了最近的策略和概念,为外肽酶和内肽酶量身定制的荧光底物库的设计和应用。
Abstract Substrate specificity of proteases can be determined using several methods among which the most frequently used are positional scanning library, proteomics and phage display. Classic approaches can deliver information about preferences for natural amino acids in binding pockets of virtually all proteases. However, recent studies demonstrate the ability to obtain much more information by application of unnatural amino acids to positional scanning library approaches. This knowledge can be used for the design of more active and specific substrates, inhibitors and activity based probes. In this minireview we describe recent strategies and concepts for the design and application of fluorogenic substrates library tailored for exopeptidases and endopeptidases.
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