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哺乳动物中的脉冲叠氮高丙氨酸标记(PALM)检测肝脏特异性LKB1基因敲除小鼠的变化。

Pulsed Azidohomoalanine Labeling in Mammals (PALM) Detects Changes in Liver-Specific LKB1 Knockout Mice.

作者信息

McClatchy Daniel B, Ma Yuanhui, Liu Chao, Stein Benjamin D, Martínez-Bartolomé Salvador, Vasquez Debbie, Hellberg Kristina, Shaw Reuben J, Yates John R

机构信息

Department of Chemical Physiology and Molecular and Cellular Molecular Biology, The Scripps Research Institute , 10550 North Torrey Pines Road, La Jolla, California 92037, United States.

Key Lab of Intelligent Information Processing, Institute of Computing Technology, Chinese Academy of Sciences , No. 6 Kexueyuan South Road, Beijing 100190, China.

出版信息

J Proteome Res. 2015 Nov 6;14(11):4815-22. doi: 10.1021/acs.jproteome.5b00653. Epub 2015 Oct 26.

Abstract

Quantification of proteomes by mass spectrometry has proven to be useful to study human pathology recapitulated in cellular or animal models of disease. Enriching and quantifying newly synthesized proteins (NSPs) at set time points by mass spectrometry has the potential to identify important early regulatory or expression changes associated with disease states or perturbations. NSP can be enriched from proteomes by employing pulsed introduction of the noncanonical amino acid, azidohomoalanine (AHA). We demonstrate that pulsed introduction of AHA in the feed of mice can label and identify NSP from multiple tissues. Furthermore, we quantitate differences in new protein expression resulting from CRE-LOX initiated knockout of LKB1 in mouse livers. Overall, the PALM strategy allows for the first time in vivo labeling of mouse tissues to differentiate protein synthesis rates at discrete time points.

摘要

通过质谱对蛋白质组进行定量已被证明有助于研究在疾病的细胞或动物模型中重现的人类病理学。通过质谱在设定时间点富集和定量新合成的蛋白质(NSP),有可能识别与疾病状态或扰动相关的重要早期调节或表达变化。通过采用非经典氨基酸叠氮高丙氨酸(AHA)的脉冲引入,可以从蛋白质组中富集NSP。我们证明,在小鼠饲料中脉冲引入AHA可以标记和鉴定来自多个组织的NSP。此外,我们定量了小鼠肝脏中由CRE-LOX启动的LKB1基因敲除导致的新蛋白质表达差异。总体而言,PALM策略首次实现了对小鼠组织的体内标记,以区分离散时间点的蛋白质合成速率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da8f/4642245/df196e546c50/pr-2015-00653c_0002.jpg

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