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）固定成糖和构成生命的蛋白质。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从周围环境中增加CO2积累的机制，这一过程被称为CO2浓缩机制（CCM）。据预测，将CCM转移到没有进化出CCM的作物上，如水稻和小麦，可能会使产量增加多达60%。有希望的CCM供体候选人是一种叫做衣藻的绿藻，它具有高效的CCM。要想成功地将藻类CCM应用到农作物中，必须知道所有的成分，它们在细胞中的位置以及它们如何共同起作用。该项目旨在快速识别构成衣藻CCM的所有蛋白质，并确定它们如何相互作用以形成增强二氧化碳吸收的功能单元。为了实现这一目标，我们将确定CCM的不同蛋白质成分在藻类细胞中的位置，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。这将使我们能够了解CCM的网络以及它是如何被监管的。最后，使用缺乏CCM单个蛋白质的突变体，我们将确定每种蛋白质成分的功能重要性。定位、相互作用和蛋白质功能数据将结合起来创建CCM的详细3D地图，可以通过开放访问的在线交互式观看平台轻松探索。预计这些数据将有助于将CCM转移到作物植物中，以增加光合作用和产量。

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.