Protein homeostasis (Proteostasis) is a description of the equilibrium state of the intracellular proteome at a specific time point, and it describes the propensity of protein molecules to be maintained at the same level in cells through the combined action of synthesis to degradation[1,2].
Plants always face various biotic and abiotic stress and research into plant proteostasis can help us better understand plant survival strategies under changing conditions. In our study, we focused on capturing plant protein synthesis. We use bio-orthogonal noncanonical amino acid tagging (BONCAT), which aims at incorporating noncanonical amino acids (ncAAs) into proteins during translation. Different from canonical amino acids, ncAAs contain active chemical groups (handles), which can react with fluorogenic dye or enrichment media for fluorescence detection or affinity enrichment, respectively[3,4]. Using BONCAT, newly synthesised proteins can be visualised or enriched, and we can better understand the rapid proteomic changes that occur in response to stress conditions.
Here, we compare some ncAAs, for their suitability for BONCAT in plants. We found that in terms of plant morphology and the viability of plant cell culture, various ncAAs performed differently. Moreover, some ncAAs may more easily implemented, for instance, the reaction of HPG with fluorogenic dye or enrichment medium requires a copper catalyst, whereas another ncAAs do not require catalysis and can react at room temperature. Thus, our study provides a systematic evaluation of different ncAAs, for nascent protein tagging in plants. Our findings will help improve the effectiveness and efficiency of BONCAT in plants.