Determination of protease cleavage site motifs using mixture-based oriented peptide libraries
Determination of protease cleavage site motifs using mixture-based oriented peptide libraries
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DOI：
10.1038/90273
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发表时间：
2001-07-01
影响因子：
46.9
通讯作者：
Cantley, LC
中科院分区：
文献类型：
--
作者：
Turk, BE;Huang, LL;Cantley, LC
作为基因组测序项目的结果，已知的蛋白酶的数量正在以惊人的速度增加。虽然人们可以通过考虑与已知蛋白酶的序列同源性来推测这些新型酶的功能，但需要新的工具来快速提供大量蛋白质的功能信息。我们描述了一种确定蛋白水解酶切割位点特异性的方法，该方法涉及多肽库混合物的混合测序。用该方法确定了基质金属蛋白酶(MMPs)家族中6种酶的切割位点基序。通过与以前文献的比较和对单独合成的多肽底物的裂解分析，验证了结果。文库数据使我们确定蛋白多糖神经聚糖是一种新的基质金属蛋白酶-2底物。我们的结果表明，一小组文库可以用来快速分析扩大的蛋白酶家族，为抑制剂的设计和蛋白质底物的鉴定提供适用的信息。
The number of known proteases is increasing at a tremendous rate as a consequence of genome sequencing projects. Although one can guess at the functions of these novel enzymes by considering sequence homology to known proteases, there is a need for new tools to rapidly provide functional information on large numbers of proteins. We describe a method for determining the cleavage site specificity of proteolytic enzymes that involves pooled sequencing of peptide library mixtures. The method was used to determine cleavage site motifs for six enzymes in the matrix metalloprotease (MMP) family. The results were validated by comparison with previous literature and by analyzing the cleavage of individually synthesized peptide substrates. The library data led us to identify the proteoglycan neurocan as a novel MMP-2 substrate. Our results indicate that a small set of libraries can be used to quickly profile an expanding protease family, providing information applicable to the design of inhibitors and to the identification of protein substrates.