Profiling deacetylase activities in cell lysates with peptide arrays and SAMDI mass spectrometry.
Profiling deacetylase activities in cell lysates with peptide arrays and SAMDI mass spectrometry.
复制标题
用肽阵列和SAMDI质谱法分析细胞裂解物中的脱乙酰基酶活性。
DOI：
10.1021/ac402614x
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发表时间：
2013-11-19
影响因子：
7.4
通讯作者：
Mrksich, Milan
中科院分区：
文献类型：
--
作者：
Kuo, Hsin-Yu;DeLuca, Teresa A.;Miller, William M.;Mrksich, Milan
能够描述细胞中分子活动的阵列的发展对于理解正常和病理环境中的信号通路是重要的。虽然寡核苷酸阵列现在被常规用于分析全球基因表达，但仍然缺乏分析细胞裂解物酶活性的工具。本文描述了自组装单分子膜上形成的多肽阵列与质谱学的结合，为鉴定细胞裂解产物中的酶活性模式提供了一种无标记的方法。该方法是通过分析CHRF巨核细胞系(Mk)细胞裂解物中的赖氨酸脱乙酰酶(KDAC)活性来证明的。类特异性脱乙酰酶抑制剂被用来显示CHRF细胞的终末MK分化的显著特征是sirtuin活性显著降低，而KDAC 1-11的活性几乎没有变化。这项工作建立了一个平台，可以用来识别各种酶活性的细胞裂解物的全局活性变化。
The development of arrays that can profile molecular activities in cells is important to understanding signaling pathways in normal and pathological settings. While oligonucleotide arrays are now routinely used to profile global gene expression, there is still a lack of tools for profiling enzyme activities in cell lysates. This paper describes the combination of peptide arrays formed on self-assembled monolayers and mass spectrometry to provide a label-free approach for identifying patterns of enzyme activities in cell lysates. The approach is demonstrated by profiling lysine deacetylase (KDAC) activities in cell lysates of the CHRF megakaryocytic (Mk) cell line. Class-specific deacetylase inhibitors were used to show that terminal Mk differentiation of CHRF cells is marked by a pronounced decrease in sirtuin activity and by little change in activity of KDACs 1-11. This work establishes a platform that can be used to identify changes in global activity profiles of cell lysates for a wide variety of enzymatic activities.
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