Monitoring peptidase activities in complex proteomes by MALDI-TOF mass spectrometry.
Monitoring peptidase activities in complex proteomes by MALDI-TOF mass spectrometry.
复制标题
通过 MALDI-TOF 质谱监测复杂蛋白质组中的肽酶活性。
DOI：
10.1038/nprot.2009.88
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发表时间：
2009
期刊：
影响因子：
14.8
通讯作者：
Tempst, Paul
中科院分区：
文献类型：
--
作者：
Villanueva, Josep;Nazarian, Arpi;Lawlor, Kevin;Tempst, Paul
Measuring enzymatic activities in biological fluids is a form of activity-based proteomics and may be utilized as a means of developing disease biomarkers. Activity-based assays allow amplification of output signals, thus potentially visualizing low-abundant enzymes on a virtually transparent whole-proteome background. The protocol presented here describes a semi-quantitative in vitro assay of proteolytic activities in complex proteomes by monitoring breakdown of designer peptide-substrates using robotic extraction and a MALDI-TOF mass spectrometric read-out. Relative quantitation of the peptide metabolites is done by comparison with spiked internal standards, followed by statistical analysis of the resulting mini-peptidome. Partial automation provides reproducibility and throughput essential for comparing large sample sets. The approach may be employed for diagnostic or predictive purposes and enables profiling of 96 samples in 30 hours. It could be tailored to many diagnostic and pharmaco-dynamic purposes, as a read-out of catalytic and metabolic activities in body fluids or tissues.
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影响因子：
7.4
作者：
Villanueva, J;Philip, J;Tempst, P
通讯作者：
Tempst, P
影响因子：
14.8
作者：
Villanueva, Josep;Lawlor, Kevin;Tempst, Paul
通讯作者：
Tempst, Paul
影响因子：
15.9
作者：
Villanueva, J;Shaffer, DR;Tempst, P
通讯作者：
Tempst, P
影响因子：
7
作者：
Villanueva, Josep;Nazarian, Arpi;Temps, Paul
通讯作者：
Temps, Paul
影响因子：
7
作者：
Villanueva, Josep;Martorella, Andrew J.;Tempst, Paul
通讯作者：
Tempst, Paul