Boonen Kurt, Husson Steven J, Landuyt Bart, Baggerman Geert, Hayakawa Eisuke, Luyten Walter H M L, Schoofs Liliane
Functional Genomics and Proteomics Research Unit, Department of Biology, K.U. Leuven, Leuven, Belgium.
Methods Mol Biol. 2010;615:191-206. doi: 10.1007/978-1-60761-535-4_15.
Endocrine tissues like the pituitary, hypothalamus and islets of Langerhans are rich in bioactive peptides. These are used for intercellular signalling and are involved in regulation of almost all physiological processes. Peptidomics is the comprehensive analysis of peptides in tissues, fluids and cells. Peptidomics applied to (neuro-)endocrine tissues aims therefore to identify as many bioactive peptides as possible. Peptidomics of (neuro-)endocrine tissues requires an integrated approach that consists of careful sample handling, peptide separation techniques, mass spectrometry and bioinformatics. Here we describe the methods for isolation and dissection of endocrine tissues, the extraction of bioactive peptides and further sample handling and identification of peptides by mass spectrometry and hyphenated techniques. We also present a straightforward method for the comparison of relative levels of bioactive peptides in these endocrine tissues under varying physiological conditions. The latter helps to elucidate functions of the bioactive peptides.
垂体、下丘脑和胰岛等内分泌组织富含生物活性肽。这些生物活性肽用于细胞间信号传导,几乎参与所有生理过程的调节。肽组学是对组织、体液和细胞中的肽进行全面分析。因此,应用于(神经)内分泌组织的肽组学旨在鉴定尽可能多的生物活性肽。(神经)内分泌组织的肽组学需要一种综合方法,该方法包括仔细的样品处理、肽分离技术、质谱分析和生物信息学。在此,我们描述了内分泌组织的分离和解剖方法、生物活性肽的提取以及进一步的样品处理,以及通过质谱分析和联用技术鉴定肽的方法。我们还提出了一种直接的方法,用于比较这些内分泌组织在不同生理条件下生物活性肽的相对水平。后者有助于阐明生物活性肽的功能。