Cell Type-specific Metabolic Labeling of Proteins with Azidonorleucine in Drosophila

Ines Erdmann; Kathrin Marter; Oliver Kobler; Sven Niehues; Julia Bussmann; Anke Müller; Julia Abele; Erik Storkebaum; Ulrich Thomas; Daniela C Dieterich

doi:10.21769/BioProtoc.2397

Cell Type-specific Metabolic Labeling of Proteins with Azidonorleucine in Drosophila

Bio Protoc. 2017 Jul 20;7(14):e2397. doi: 10.21769/BioProtoc.2397.

Authors

Ines Erdmann^{1

2}, Kathrin Marter^{1

2

3}, Oliver Kobler⁴, Sven Niehues^{5

6}, Julia Bussmann^{5

6}, Anke Müller^{1

2}, Julia Abele^{1

2}, Erik Storkebaum^{5

6}, Ulrich Thomas⁴, Daniela C Dieterich^{1

2

3}

Affiliations

¹ Neuronal Plasticity and Communication, Institute for Pharmacology and Toxicology, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany.
² Research Group Neuralomics, Leibniz Institute for Neurobiology, Magdeburg, Germany.
³ Center for Behavioral Brain Sciences, Magdeburg, Germany.
⁴ Department of Neurochemistry and Molecular Biology, Leibniz Institute for Neurobiology, Magdeburg, Germany.
⁵ Molecular Neurogenetics Laboratory, Max Planck Institute for Molecular Biomedicine, Münster, Germany.
⁶ Faculty of Medicine, University of Münster, Münster, Germany.

Abstract

Advanced mass spectrometry technology has pushed proteomic analyses to the forefront of biological and biomedical research. Limitations of proteomic approaches now often remain with sample preparations rather than with the sensitivity of protein detection. However, deciphering proteomes and their context-dependent dynamics in subgroups of tissue-embedded cells still poses a challenge, which we meet with a detailed version of our recently established protocol for cell-selective and temporally controllable metabolic labeling of proteins in Drosophila. This method is based on targeted expression of a mutated variant of methionyl-tRNA-synthetase, MetRS^L262G, which allows for charging methionine tRNAs with the non-canonical amino acid azidonorleucine (ANL) and, thus, for detectable ANL incorporation into nascent polypeptide chains.

Keywords: Click chemistry; Drosophila melanogaster; Metabolic labeling; Protein synthesis; Proteomic profiling.