Recently, Dr. Chen Yanmei, Key Laboratory of plant physiology and biochemistry, China Agricultural University, published a review paper entitled mass specialty untangles plant membrane protein signaling networks at trends in plant science. Since its invention, mass spectrometry has won many Nobel prizes and has been widely used in plant biology research in recent years. This review summarizes the latest progress in the study of plant cell membrane proteins, especially the function of kinases, membrane protein phosphorylation and protein interaction by using mass spectrometry. It comprehensively expounds the analytical thinking of plant membrane signal network, and discusses the application of these technologies in plant biology, and puts forward effective analytical methods.
As an important component of material transport, energy conversion and signal sensing between plants and their environment, cytoplasmic membrane plays an important role in the life process of plant growth and development and response to adversity. Membrane proteins are the main carriers of plasma membrane function. According to their distribution and location on plasma membrane, they are mainly divided into three categories: outer membrane proteins, integrated membrane proteins and lipid anchored proteins. The plasma membrane is rich in receptor kinases, channel proteins and enzymes, which play a key role in plant physiological and biochemical processes. However, it is very difficult to analyze and detect membrane protein because of its high complexity, low relative content and strong hydrophobicity. On the basis of these research areas in recent years, this review focuses on the application of mass spectrometry in the study of plasma membrane signal network, including the full spectrum analysis of plasma membrane protein, plasma membrane protein phosphorylation, protein kinase substrate interaction, lipid raft microdomain protein and membrane protein interaction.
Figure 1. Research method of plasma membrane protein signal network based on mass spectrometry
It is very difficult to detect membrane protein and its phosphorylation, and it is easy to be interfered by background noise. In this review, the technical challenges and solutions for the study and analysis of membrane proteins and phosphorylation modification of membrane proteins are described. In addition, the enrichment and purification methods of protein phosphorylation, as well as the problems needing attention in practical application were discussed.
Fig. 2. Signal network of plasma membrane protein in plant cells.
In addition, the application of mass spectrometry in the study of proteomics and phosphorylation of lipid rafts was reviewed. In addition to the summary of qualitative and quantitative correlation analysis methods of this kind of protein, this paper focuses on the relevant methods and applications of mass spectrometry on how to mine the target substrate of kinase. On the basis of these, the research progress of mass spectrometry in the field of membrane protein interaction is introduced in detail. At the same time, how to study the interaction of phosphorylated regulatory proteins by mass spectrometry is also discussed, which provides a new idea for the research of plant plasma membrane proteins. Finally, it is pointed out that the high-throughput analysis of cytoplasmic membrane protein interaction by mass spectrometry remains to be explored, and there is still a gap in the study of the relationship between multiple post-translational modifications of proteins.
As the first unit of this paper, Dr. Chen Yanmei and Prof. wolfram Weckwerth, center of metabolomics, University of Vienna, Austria, are co authors. This paper is supported by NSFC.
Paper link: https://doi.org/10.1016/j.tplants.2020.03.013