Molecular characterisation of an ADP-dependent regulatory protein

ADP 依赖性调节蛋白的分子表征

• 批准号: BB/G004145/1
• 负责人: Michael Webb
• 金额: $ 41.62万
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2008
• 资助国家: 英国
• 起止时间: 2008 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FG004145%2F1
• 关键词: Molecular; characterisation; ADP; dependent; regulatory

Conversion of carbon dioxide to sugars by plants is essential to life on earth. In the majority of plants this process competes with non-productive reaction with oxygen. One group of plants, the C4 plants, have developed a strategy to avoid this: this strategy allows the plants to survive in hot, dry and arid conditions which would not otherwise be viable for plant growth. These plants include the common crop plants, maize and sugarcane. Research is presently underway to determine if this pathway can be engineered into other plants to generate hardier crop plants and this research will feed into this objective. Our research addresses how one part of this alternative strategy of making sugars is controlled. In particular, we are interested in how an unusual protein acts to switch off the pathway in the dark. The protein, pyruvate,orthophosphate dikinase (PPDK) regulatory protein, or PDRP for short, inactivates another protein, PPDK, thereby preventing the plant from making the compound phosphoenolpyruvate from pyruvate. This prevents the plant from 'fixing' carbon dioxide into sugars in the dark (when there is no light to provide energy via photosynthesis). The regulatory protein, PDRP, can perform this vital function because it is dependent upon the presence of its substrate, ADP, for activity. This accumulates in the dark as a direct result of photosynthesis shutting down. We are interested in the details of the way in which the regulatory protein uses this molecule to inactivate PPDK. We will develop new ways to study this process and make mimics of the target which will help us further unravel the precise manner in which it occurs. The target protein is inactivated because a phosphate group is transferred from the substrate ADP to the protein, thereby blocking the part of PPDK required to perform its normal function. It is worth noting that using this particular molecule, adenine diphosphate (ADP), to perform this reaction is highly unusual, normally adenine triphosphate (ATP) is used instead.

植物将二氧化碳转化为糖对地球上的生命至关重要。在大多数工厂中，这个过程与氧气的非生产性反应竞争。一组植物，C4植物，已经开发出一种策略来避免这种情况：这种策略允许植物在炎热，干燥和干旱的条件下生存，否则植物生长将无法生存。这些植物包括常见的作物，玉米和甘蔗。目前正在进行研究，以确定是否可以将这种途径工程化到其他植物中，以产生更耐寒的作物植物，这项研究将有助于实现这一目标。我们的研究解决了如何控制这种制造糖的替代策略的一部分。特别是，我们感兴趣的是一种不寻常的蛋白质如何在黑暗中关闭通路。丙酮酸正磷酸二激酶（PPDK）调节蛋白（简称PDRP）使另一种蛋白质PPDK失活，从而阻止植物从丙酮酸合成磷酸烯醇丙酮酸。这可以防止植物在黑暗中（当没有光通过光合作用提供能量时）将二氧化碳“固定”为糖。调节蛋白PDRP可以执行这一重要功能，因为它依赖于其底物ADP的存在来进行活性。这是光合作用停止的直接结果。我们感兴趣的是调节蛋白使用这种分子来抑制PPDK的方式的细节。我们将开发新的方法来研究这一过程，并模仿目标，这将有助于我们进一步揭示它发生的精确方式。靶蛋白失活是因为磷酸基团从底物ADP转移到蛋白质上，从而阻断了PPDK执行其正常功能所需的部分。值得注意的是，使用这种特殊的分子腺嘌呤二磷酸（ADP）来进行这种反应是非常不寻常的，通常使用腺嘌呤三磷酸（ATP）代替。

项目成果

Confirmation of a Protein-Protein Interaction in the Pantothenate Biosynthetic Pathway by Using Sortase-Mediated Labelling.

• DOI: 10.1002/cbic.201500547
• 发表时间: 2016-04-15
• 期刊: Chembiochem : a European journal of chemical biology
• 作者: Morrison PM;Balmforth MR;Ness SW;Williamson DJ;Rugen MD;Turnbull WB;Webb ME
• 通讯作者: Webb ME

Efficient N-terminal labeling of proteins by use of sortase.

• DOI: 10.1002/anie.201204538
• 发表时间: 2012-09-10
• 期刊: Angewandte Chemie (International ed. in English)
• 作者: Williamson, Daniel J;Fascione, Martin A;Turnbull, W Bruce
• 通讯作者: Turnbull, W Bruce

Prospects for stable analogues of phosphohistidine.

• DOI: 10.1042/bst20130071
• 发表时间: 2013-08
• 期刊: Biochemical Society transactions
• 影响因子: 3.9
• 作者: Tom E. McAllister;J. Hollins;M. Webb