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Dithiothreitol (DTT)


Company: VWR
Catalog#: V3155
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Preparation and Analysis of Crude Autolytic Enzyme Extracts from Staphylococcus aureus
[Abstract]  The metabolism of the cell surface during bacterial cell division involves synthesis and degradation of peptidoglycan (PGN), the major component of the bacterial cell wall. Bacteria have to ensure that their surface remains capable of withstanding high turgor pressures and, simultaneously, that the PGN at their surface is concealed from receptors produced by the host innate immune system. For cell separation to occur, and for PGN to be kept concealed, “old” PGN is degraded by specific PGN hydrolases, also known as autolysins, that are found at the bacterial cell surface or that are secreted into the growth medium.

Bacterial PGN hydrolases are cell wall lytic enzymes that comprise a broad and diverse group of proteins. It is often difficult to assign a specific function to a ...
[摘要]  细菌细胞分裂期间细胞表面的代谢涉及细菌细胞壁的主要组分肽聚糖(PGN)的合成和降解。细菌必须确保它们的表面能够承受高的膨胀压力,同时,它们表面的PGN被宿主先天免疫系统产生的受体所掩盖。为了发生细胞分离,并且对于PGN保持隐蔽,“老”PGN由在细菌细胞表面发现或分泌到生长培养基中的特定PGN水解酶(也称为自溶素)降解。
细菌PGN水解酶是包含广泛和多样化蛋白质组的细胞壁溶解酶。主要是因为生物体可以具有大量具有冗余活性的水解酶,并且一种水解酶可以具有多种酶活性并参与各种细胞过程(Vollmer等,2008),通常难以将特定功能分配给PGN水解酶。 。枯草芽孢杆菌35种已知或假设的PGN水解酶,而大肠杆菌(E.coli)和金黄色葡萄球菌(金黄色葡萄球菌)分别具有约16和19个PGN水解酶(Vollmer,2012; Heidrich等,2001; Singh et al。 ,2012)。
在这里,我们描述了提取与金黄色葡萄球菌细胞壁非共价连接的PGN水解酶的方法(Vollmer,2008)。含有变性PGN水解酶的提取物的分析是通过运行Zymogram凝胶(含有粗细菌细胞壁或底物细胞的SDS-PAGE凝胶)进行的,然后将其在非变性缓冲液中温育以允许PGN水解酶复性。然后可以通过产生在细胞壁消化发生时观察到的清除条带来鉴定这些复性酶。方案分为三个步骤:A)从金黄色葡萄球菌细胞中制备粗自动提取物; ...

A Phosphopeptide Purification Protocol for the Moss Physcomitrella paten
[Abstract]  Protein phosphorylation is one of the most common post-translational modifications in eukaryotic cells and plays a critical role in a vast array of cellular processes. Efficient methods of protein extraction and phosphopeptide purification are required to ensure the detection of high quality of proteins. In our hands, phenol extraction of proteins and TiO2 chromatography enrich phosphorylated peptides more efficiently than other methods in the moss Physcomitrlla patens (P. patens). [摘要]  蛋白磷酸化是真核细胞中最常见的翻译后修饰之一,并在大量细胞过程中起关键作用。 需要蛋白质提取和磷酸肽纯化的有效方法以确保高质量的蛋白质的检测。 在我们的手中,蛋白质和TiO 2层析的酚提取比在小麦Physcomitrlla patens中的其它方法更有效地富集磷酸化肽。 )。