Karpiński Adam Aleksander, Torres Elguera Julio Cesar, Sanner Anne, Konopka Witold, Kaczmarek Leszek, Winter Dominic, Konopka Anna, Bulska Ewa
Faculty of Chemistry, University of Warsaw, Pasteura 1, 02-093 Warsaw, Poland.
Nencki Institute of Experimental Biology of the Polish Academy of Sciences, Pasteura 3, 02-093 Warsaw, Poland.
Biomedicines. 2022 Oct 2;10(10):2466. doi: 10.3390/biomedicines10102466.
Mass spectrometry-based proteomics aims to study the proteome both qualitatively and quantitatively. A key step in proteomic analysis is sample preparation, which is crucial for reliable results. We investigated the effect of the composition of the homogenization buffer used to extract proteins from brain tissue on the yield of protein extraction and the number and type of extracted proteins. Three different types of buffers were compared-detergent-based buffer (DB), chaotropic agent-based buffer (CAB) and buffer without detergent and chaotropic agent (DFB). Based on label-free quantitative protein analysis, detergent buffer was identified as the most suitable for global proteomic profiling of brain tissue. It allows the most efficient extraction of membrane proteins, synaptic and synaptic membrane proteins along with ribosomal, mitochondrial and myelin sheath proteins, which are of particular interest in the field of neurodegenerative disorders research.
基于质谱的蛋白质组学旨在对蛋白质组进行定性和定量研究。蛋白质组分析的关键步骤是样品制备，这对于获得可靠结果至关重要。我们研究了用于从脑组织中提取蛋白质的匀浆缓冲液组成对蛋白质提取产量以及提取的蛋白质数量和类型的影响。比较了三种不同类型的缓冲液——基于去污剂的缓冲液（DB）、基于离液剂的缓冲液（CAB）和不含去污剂和离液剂的缓冲液（DFB）。基于无标记定量蛋白质分析，去污剂缓冲液被确定为最适合脑组织整体蛋白质组分析的缓冲液。它能够最有效地提取膜蛋白、突触和突触膜蛋白以及核糖体、线粒体和髓鞘蛋白，这些蛋白在神经退行性疾病研究领域具有特别的研究意义。
