Fluorous-based peptide microarrays for protease screening
Fluorous-based peptide microarrays for protease screening
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DOI：
10.1016/j.jfluchem.2009.09.005
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发表时间：
2009-11-01
影响因子：
1.9
通讯作者：
Pohl, Nicola L. B.
中科院分区：
文献类型：
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作者：
Collet, Beatrice Y. M.;Nagashima, Tadamichi;Pohl, Nicola L. B.
随着越来越多的蛋白酶序列通过基因组测序项目被发现，有效的平行方法来发现这些蛋白酶的潜在底物变得至关重要。在这里，我们描述了首次使用基于氟的微阵列来探测肽序列，并开始定义用于蛋白酶筛选的氟微阵列技术的范围和局限性。一系列丝氨酸蛋白酶的比较表明，将这些底物固定在含氟玻片上后，它们在溶液中切割肽底物的能力得以维持。含氟表面不能显著灭活酶。然而，在多肽序列中加入亲水组分可以诱导与疏水部分有亲和关系的酶（如凝乳胰蛋白酶）对底物的切割速率降低。这项工作代表了使用非共价微阵列接口创建强大的蛋白酶筛选平台的第一步，该平台可以很容易地在同一载玻片上合并一系列化合物。(C) 2009 Elsevier B.V.版权所有
As ever more protease sequences are uncovered through genome sequencing projects, efficient parallel methods to discover the potential substrates of these proteases becomes crucial. Herein we describe the first use of fluorous-based microarrays to probe peptide sequences and begin to define the scope and limitations of fluorous microarray technologies for the screening of proteases. Comparison of a series of serine proteases showed that their ability to cleave peptide substrates in solution was maintained upon immobilization of these substrates onto fluorous-coated glass slides. The fluorous surface did not serve to significantly inactivate the enzymes. However, addition of hydrophilic components to the peptide sequences could induce lower rates of substrate cleavage with enzymes such as chymotrypsin with affinities to hydrophobic moieties. This work represents the first step to creating robust protease screening platforms using noncovalent microarray interface that can easily incorporate a range of compounds on the same slide. (C) 2009 Elsevier B.V. All rights reserved.