Experimental setup for the identification of mitochondrial protease substrates by shotgun and top-down proteomics

Alice Di Pierro; Heather Bondi; Chiara Monti; Luisa Pieroni; Enrico Cilio; Andrea Urbani; Tiziana Alberio; Mauro Fasano; Maurizio Ronci

doi:10.1016/j.euprot.2016.02.002

Experimental setup for the identification of mitochondrial protease substrates by shotgun and top-down proteomics

EuPA Open Proteom. 2016 Feb 22:11:1-3. doi: 10.1016/j.euprot.2016.02.002. eCollection 2016 Jun.

Authors

Alice Di Pierro¹, Heather Bondi^{1

2}, Chiara Monti^{1

2}, Luisa Pieroni^{3

4}, Enrico Cilio^{3

4}, Andrea Urbani^{3

4}, Tiziana Alberio^{1

2}, Mauro Fasano^{1

2}, Maurizio Ronci⁵

Affiliations

¹ Department of Science and High Technology, University of Insubria, I-21052 Busto Arsizio, Italy.
² Center of Neuroscience, University of Insubria, I-21052 Busto Arsizio, Italy.
³ Santa Lucia IRCCS Foundation, I-00143 Rome, Italy.
⁴ Department of Experimental Medicine and Surgery, University of Rome "Tor Vergata", I-00133 Rome, Italy.
⁵ Dept. of Medical, Oral and Biotechnological Sciences, University "G. D'Annunzio" of Chieti-Pescara, I-66013 Chieti, Italy.

Abstract

Mitochondria possess a proteolytic system that contributes to the regulation of mitochondrial dynamics, mitochondrial biogenesis and mitophagy. We aimed at the identification by bottom-up proteomics of altered protein processing due to the activation of mitochondrial proteases in a cellular model of impaired dopamine homeostasis. Moreover, we optimized the conditions for top-down proteomics to identify the cleavage site sequences.

Keywords: Electroelution; Mitochondria; Protease; Shotgun; Top-down.