Oral Presentation HUPO 2019 - 18th Human Proteome Organization World Congress

Is the mitochondrial protein processing system robust? Lessons from a combined N-terminomics and shotgun proteomics approach on human cells treated with rapamycin or zinc (#92)

Thierry Rabilloud 1 , Joanna Bons 2 , Charlotte Macron 2 , Catherine Aude-Garcia 1 , Sebastian Alvaro Vaca-Jacome 2 , Magali Rompais 2 , Sarah Cianferani 2 , Christine Carapito 2
  1. Chemistry and Biology of Metals, UMR CNRS-CEA-UGA 5249, Grenoble, RA, France
  2. IPHC/LSMBP, Strasbourg, Alsace, France

All but thirteen mammalian mitochondrial proteins are encoded by the nuclear genome, translated in the cytosol and then imported into the mitochondria. For a significant proportion of the mitochondrial proteins, import is coupled with the cleavage of a presequence called the transit peptide, and the formation of a new N-terminus. Determination of the neo N-termini has been investigated by proteomic approaches in several systems, but generally in a static way in order to compile as many N-termini as possible. In the present study, we have investigated how the mitochondrial proteome and N-terminome react to chemical stimuli that alter mitochondrial metabolism, namely zinc ions and rapamycin. To this end, we have used a strategy that analyzes both internal and N-terminal peptides in a single run, the dN-TOP approach. Rapamycin and zinc induced different changes in the mitochondrial proteome. However, convergent changes to key mitochondrial enzymatic activities such as pyruvate dehydrogenase, succinate dehydrogenase and citrate synthase were observed for both treatments. Other convergent changes were seen in components of the N-terminal processing system and mitochondrial proteases. Investigations into the generation of neo N-termini in mitochondria showed that the processing system is robust, as indicated by the lack of change in neo N-termini under the conditions tested. Detailed analysis of the data revealed that zinc caused a slight reduction in the efficiency of the N-terminal trimming system and that both treatments increased the degradation of mitochondrial proteins.

In conclusion, the use of this combined strategy allowed a detailed analysis of the dynamics of the mitochondrial N-terminome in response to treatments which impact the mitochondria.