Analysis of the dynamic sulfotyrosine proteome.

生物体的生存取决于不同类型的细胞通过在正确的时间和正确的地点组装正确的蛋白质复合体来相互沟通的能力。实现这一目标的一种方法是使用化学和生物催化剂(酶)的技巧，称为酪氨酰蛋白磺基转移酶，或TPST，以改变聚合物的生物特性，如蛋白质。作为一种快速(通常是)可逆调节的手段，TPST通过添加少量带电化学品来推动这一过程。这些事件，更准确地说，被称为‘翻译后修饰’，或PTM，作为开关来改变信息流，并决定所引发的不同生物结果的类型，如细胞运动、生长、存活或死亡。我们的建议旨在利用新的工具和蛋白质组学技术(使用质谱学，以公正的方式观察细胞中的所有蛋白质)来评估在蛋白质中添加一种特定的化学基团，称为硫酸盐。目前，对硫酸盐的分析没有重点，吸引的战略资金很少，而且很难进行全面的操作或研究，这使得努力研究其全球意义具有挑战性。由于它支撑了如此多的基础生物学，但关于这一调控机制的机制的详细信息仍然缺乏，硫酸盐化研究需要协调一致的战略，以制定分析、汇编和分发与蛋白质硫酸盐如何调控细胞和非细胞生物学有关的大量生物数据的方法。事实上，以技术为基础的方法分析蛋白质上的另一种化学基团磷酸，导致了我们对细胞通信方式的理解的一场革命，40多年来，对于在结构生物学、细胞信号和通信以及药物设计领域工作的生物学家来说，这一方法至关重要，并对世界各地的生物技术、制药工业和临床干预产生了连锁影响。为了快速推进我们对细胞蛋白酪氨酸硫化(STYR)的理解，我们的目标将通过相关但不同的工作包来实现。它们是：WP1：多种多肽和蛋白质中STYR(和STHR/SSER)位点特异性的生化分析WP2：从复杂细胞来源的混合物中分离STYR和基于质谱学的定量STYR分析WP3：优化基于细胞的方法来操纵哺乳动物细胞中的TPST1/2和干扰STYR含量WP4：质谱学数据的计算分析和用于公共分布和生物推断的酪氨酸硫化网络的解剖WP5：分析与酪氨酸硫化相关的新细胞角色的工作流程

The survival of an organism depends upon the ability of different cell types to communicate with each other by assembling the correct complexes of proteins at the correct time in the correct place. One way this is achieved is to use the tricks of chemistry and biological catalysts (enzymes) called tyrosyl protein sulfotransferases, or TPSTs, to change the biological properties of polymers, such as proteins. TPSTs drive this process by adding small charged chemicals as a means of rapid (usually) reversible regulation. These events, more accurately called 'post-translational modifications', or PTMs, act as switches to change information flow and dictate the types of different biological outcomes elicited, such as cell movement, growth, survival or death. Our proposal aims to exploit new tools and proteomics technology (using mass spectrometry, which looks at all proteins in cells in an unbiased way) to evaluate the addition of a specific chemical group, called sulfate, to proteins. Currently, the analysis of sulfation is unfocused, it attracts little strategic funding, and it is difficult to manipulate or study in a holistic manner, making efforts to study its global significance challenging. Since it underpins so much basic biology, yet detailed information as to the mechanism of this regulatory mechanism is still lacking, sulfation research requires concerted strategies to develop ways of analysing, compiling and distributing, the large amounts of biological data pertaining to how protein sulphation regulates cellular and acellular biology. Indeed, technology-based approaches for the analysis of a different chemical group on proteins, phosphate, led to a revolution in our understanding of how cells communicate, and has been critically important for biologists working in the areas of structural biology, cell signalling and communication and drug design for over 40 years, with knock-on effects on biotechnology, pharmaceutical industries and clinical intervention across the world. To rapidly advance our understanding of cellular protein tyrosine sulfation (sTyr), our aims will be achieved through related, but distinct, work packages. These are:WP1: Biochemical analysis of sTyr (and sThr/sSer) site-specificity in a variety of peptides and proteins WP2: sTyr isolation and mass spectrometry-based quantitative sTyr analysis from complex cell-derived mixturesWP3: Optimise cell-based approaches to manipulate TPST1/2 and perturb sTyr content in mammalian cellsWP4: Computational analysis of mass spectrometry data and dissection of tyrosine sulfation networks for public distribution and biological inferenceWP5: A workflow for analysis of new cellular roles associated with Tyr sulphation