Profiling the specific reactivity of the proteome with non-directed activity-based probes
Profiling the specific reactivity of the proteome with non-directed activity-based probes
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DOI：
10.1016/s1074-5521(00)90060-7
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发表时间：
2001-01-01
影响因子：
--
通讯作者：
Sorensen, EJ
中科院分区：
文献类型：
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作者：
Adam, GC;Cravatt, BF;Sorensen, EJ
背景：蛋白质组学领域的目标是在全球水平上描述蛋白质功能的动态变化。然而，几类蛋白质，特别是低丰度蛋白质，仍然很难用标准的蛋白质组学技术来表征。最近，出现了基于活性而不是数量的蛋白质分类的化学策略，从而极大地促进了蛋白质组低丰度成分的分析。结果：为了扩大基于活性的方法容易分析的蛋白质类别，我们合成了一个生物素化磺酸酯文库，并将其成员应用到复杂蛋白质组中，在区分特定蛋白质反应性模式的条件下。个别磺酸盐表现出独特的蛋白质组反应性，在极端情况下，它们几乎相互垂直。一个强健标记的蛋白质被鉴定为I类乙醛脱氢酶，并被磺酸盐文库成员不可逆地抑制，结论：通过非定向化学探针文库筛选蛋白质组，发现了不同的蛋白质反应模式。这些探针基于丰度以外的性质标记蛋白质靶标，绕过了当代蛋白质组学研究面临的主要挑战之一。进一步考虑到发现这些探针抑制了目标酶的催化活性。这里描述的方法应该有助于鉴定具有选择性蛋白质组反应性和新的生物活性的化合物。(C)2001爱思唯尔科学有限公司。保留所有权利。
Background: The field of proteomics aims to characterize dynamics in protein function on a global level. However, several classes of proteins, in particular low abundance proteins, remain difficult to characterize using standard proteomics technologies. Recently, chemical strategies have emerged that profile classes of proteins based on activity rather than quantity, thereby greatly facilitating the analysis of low abundance constituents of the proteome.Results: In order to expand the classes of proteins susceptible to analysis by activity-based methods, we have synthesized a library of biotinylated sulfonate esters and applied its members to complex proteomes under conditions that distinguish patterns of specific protein reactivity. Individual sulfonates exhibited unique profiles of proteome reactivity that in extreme cases appeared nearly orthogonal to one another. A robustly labeled protein was identified as a class I aldehyde dehydrogenase and shown to be irreversibly inhibited by members of the sulfonate library,Conclusions: Through screening the proteome with a nondirected library of chemical probes, diverse patterns of protein reactivity were uncovered. These probes labeled protein targets based on properties other than abundance, circumventing one of the major challenges facing contemporary proteomics research. Considering further that the probes were found to inhibit a target enzyme's catalytic activity. the methods described herein should facilitate the identification of compounds possessing both selective proteome reactivities and novel bioactivities. (C) 2001 Elsevier Science Ltd. All rights reserved.