Secondary‐Ion Mass Spectrometry of Genetically Encoded Targets |
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| Authors: | Ingrid C Vreja Selda Kabatas Dr Sinem K Saka Katharina Kröhnert Dr Carmen Höschen Dr Felipe Opazo Prof?Dr Ulf Diederichsen Prof?Dr Silvio O Rizzoli |
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| Institution: | 1. Department of Neuro‐ and Sensory Physiology, University Medical Center G?ttingen, Humboldtallee 23, 37073 G?ttingen (Germany);2. Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), G?ttingen (Germany);3. International Max Planck Research School Molecular Biology, G?ttingen (Germany);4. Institute for Organic and Biomolecular Chemistry, University of G?ttingen, Tammannstrasse 2, 37077 G?ttingen (Germany);5. Department of Ecology and Ecosystem Management, Center of Life and Food Sciences Weihenstephan, Technische Universit?t München, Freising‐Weihenstephan (Germany) |
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| Abstract: | Secondary ion mass spectrometry (SIMS) is generally used in imaging the isotopic composition of various materials. It is becoming increasingly popular in biology, especially for investigations of cellular metabolism. However, individual proteins are difficult to identify in SIMS, which limits the ability of this technology to study individual compartments or protein complexes. We present a method for specific protein isotopic and fluorescence labeling (SPILL), based on a novel click reaction with isotopic probes. Using this method, we added 19F‐enriched labels to different proteins, and visualized them by NanoSIMS and fluorescence microscopy. The 19F signal allowed the precise visualization of the protein of interest, with minimal background, and enabled correlative studies of protein distribution and cellular metabolism or composition. SPILL can be applied to biological systems suitable for click chemistry, which include most cell‐culture systems, as well as small model organisms. |
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| Keywords: | click chemistry isotopic labeling protein engineering secondary‐ion mass spectrometry (SIMS) unnatural amino acid |
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