Creating a spatially defined, multidimensional, protein interactome of the eukaryotic algal CO2 concentrating mechanism

光合作用利用来自太阳的能量将二氧化碳(CO2)固定为糖和蛋白质，构成生命的组成部分。为了增强光合作用，一些植物和几乎所有的藻类都进化出了增加周围环境中二氧化碳积累的机制，这个过程被称为二氧化碳浓缩机制(CCM)。据预测，将CCM转移到水稻和小麦等未能进化出CCM的作物上，可以增加高达60%的产量。一种有希望的CCM捐赠者是一种名为衣藻的绿藻，它具有高效的CCM。为了成功地在作物中实施藻类CCM，至关重要的是所有的成分都是已知的，它们在细胞中的位置以及它们是如何共同发挥作用的。该项目旨在快速确定组成衣藻CCM的所有蛋白质，并确定它们如何相互作用，形成一个增强二氧化碳吸收的功能单位。为了实现这一点，我们将确定CCM的不同蛋白质组分在藻细胞中的位置，然后我们将确定它们与哪些蛋白质相互作用，以及当CCM打开和关闭时，这些相互作用如何变化。这将使我们了解CCM的网络以及它是如何受到监管的。最后，使用缺乏CCM个别蛋白质的突变体，我们将确定每个蛋白质组分的功能重要性。定位、相互作用和蛋白质功能数据将被结合起来，以创建CCM的详细3D地图，该地图可以通过开放访问的在线互动查看平台轻松探索。预计这些数据将有助于将CCM转移到农作物中，以提高光合作用和产量。

Photosynthesis harnesses energy from the sun to fix carbon dioxide (CO2) into sugars and the protein building blocks of life. To enhance photosynthesis some plants and nearly all algae have evolved mechanisms to increase the accumulation of CO2 from their surrounding environment, this process in known as a CO2 concentrating mechanism (CCM). It is predicted that the transfer of a CCM to crop plants, such as rice and wheat that have failed to evolve CCMs, could increase yields by up to 60%. A promising CCM donor candidate is a green alga called Chlamydomonas, which has a highly efficient CCM. For the successful implementation of an algal CCM into crop plants it is essential that all the components are known, where they are found in the cell and how they function together. This project aims to rapidly identify all the proteins that make up the Chlamydomonas CCM and to determine how they interact with each other to form a functional unit that enhances CO2 uptake. To achieve this, we will determine where the different protein components of the CCM are located in the algal cell, we will then identify what proteins they are interacting with and how these interactions change when the CCM is switched on and off. This will allow us to understand the network of the CCM and how it is regulated. Finally, using mutants that lack individual proteins of the CCM, we will determine the functional importance of each protein component. The localisation, interaction and protein function data will be combined to create a detailed 3D map of the CCM that can be easily explored through an open-access, online interactive viewing platform. It is anticipated that these data will facilitate the transfer of a CCM into crop plants to increase photosynthesis and yields.