A highly efficient method for extracting peptides from a single mouse hypothalamus
A highly efficient method for extracting peptides from a single mouse hypothalamus
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DOI：
10.1016/j.bbrc.2021.02.041
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发表时间：
2021-02-25
影响因子：
3.1
通讯作者：
Kodera, Yoshio
中科院分区：
文献类型：
--
作者：
Nakagawa, Yuzuru;Matsui, Takashi;Kodera, Yoshio
生物体内含有多种内源性多肽，它们对许多生物过程起着重要的调节作用。内源性多肽通常使用液质联用(LC-MS)进行分析。然而，由于多肽提取效率低，单个动物的多肽丰度低，基于LC-MS的多肽组学研究并没有促进对多肽丰度的个体差异和组织特异性的了解。在本研究中，我们建立了一种多肽的提取方法，并随后进行了纳米流动LC-MS/MS分析。这种方法使得从单个动物的亚毫克量的下丘脑中提取多肽变得高效和可重复性。从相当于135微克下丘脑的样本体积中检测到各种生物活性和正品多肽。这种方法可能有助于阐明个体差异和组织特异性，也有助于发现新的生物活性多肽和生物标记物，并开发多肽治疗药物。(C)2021 Elsevier Inc.保留所有权利。
Living organisms contain a variety of endogenous peptides that function as significant regulators of many biological processes. Endogenous peptides are typically analyzed using liquid chromatographyemass spectrometry (LC-MS). However, due to the low efficiency of peptide extraction and low abundance of peptides in a single animal, LC-MSebased peptidomics studies have not facilitated an understanding of the individual differences and tissue specificity of peptide abundance. In this study, we developed a peptide extraction method followed by nano-flow LC-MS/MS analysis. This method enabled highly efficient and reproducible peptide extraction from sub-milligram quantities of hypothalamus dissected from a single animal. Diverse bioactive and authentic peptides were detected from a sample volume equivalent to 135 mu g of hypothalamus. This method may be useful for elucidating individual differences and tissue specificity, as well as for facilitating the discovery of novel bioactive peptides and biomarkers and developing peptide therapeutics. (C) 2021 Elsevier Inc. All rights reserved.