Secondary-ion mass spectrometry of genetically encoded targets
| Autor: | Sinem K. Saka, Katharina Kröhnert, Ulf Diederichsen, Carmen Höschen, Silvio O. Rizzoli, Ingrid C. Vreja, Felipe Opazo, Selda Kabatas |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Fluorine Radioisotopes
Analytical chemistry Spectrometry Mass Secondary Ion 010402 general chemistry 01 natural sciences Catalysis Cell Line Isotopic labeling 03 medical and health sciences Cricetinae Stable isotope labeling by amino acids in cell culture Fluorescence microscope Animals Nanotechnology unnatural amino acid click chemistry isotopic labeling protein engineering secondary-ion mass spectrometry (SIMS) Fluorescent Dyes 030304 developmental biology 0303 health sciences Cellular metabolism Molecular Structure Chemistry Proteins General Chemistry Protein engineering Fluorescence Communications 0104 chemical sciences Secondary ion mass spectrometry Microscopy Fluorescence Click chemistry Biophysics |
| Zdroj: | Angewandte Chemie (International Ed. in English) |
| Popis: | 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 (19) F-enriched labels to different proteins, and visualized them by NanoSIMS and fluorescence microscopy. The (19) F 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. peerReviewed |
| Databáze: | OpenAIRE |
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